JP2019505568A - Bicyclic inhibitor of PAD4 - Google Patents

Abstract

The present invention provides compounds useful as inhibitors of PAD4, compositions thereof and methods of treating PAD4-related diseases.

Description

(Background of the invention)
PAD4 is a member of the peptidylarginine deiminase (PAD) family of enzymes capable of catalyzing citrullination of arginine to citrulline within the peptide sequence. PAD4 has been implicated in the deimination or citrullination of various proteins in vitro and in vivo resulting in diverse functional responses in various diseases (Jones JE et al., Curr. Opin. Drug Discov. Devel ., 12 (5), (2009), 616-627). Examples of exemplary diseases include, besides oncological indications, rheumatoid arthritis, diseases involving neutrophils in etiology (e.g. vasculitis, systemic lupus erythematosus, ulcerative colitis) . PAD4 inhibitors may also have broader applicability as tools and treatments for diseases with human epigenetic mechanisms.

  Inhibitors of PAD4 have utility against rheumatoid arthritis (RA). RA is an autoimmune disease that affects approximately 1% of the population (Wegner N et al., Immunol. Rev., 233 (1) (2010), 34-54). RA is characterized by inflammation of the articular joints leading to a weakened destruction of bone and cartilage. Several population studies have suggested a weak genetic association between PAD4 polymorphisms and susceptibility to RA, although there are inconsistencies (Kochi Y. et al., Ann. Rheum. Dis. , 70, (2011), 512-515). PAD4 (along with the family member PAD2) has been detected in synovial tissue involved in the deimination of various joint proteins. This process is presumed to lead to a breakthrough of tolerance and initiation of immune responses to citrullinated substrates such as fibrinogen, vimentin and collagen in RA joints. These anti-citrullinated protein antibodies (ACPA) contribute to the pathogenesis of the disease and can also be used as a diagnostic test for RA (for example, commercially available CCP2 or cyclic citrullination) Protein 2 reagent). Furthermore, increased citrullination leads to direct involvement in further disease pathogenesis through its ability to directly affect some joint functions and inflammatory mediators (e.g., fibrinogen, antithrombin, multiple chemokines) There is also a fear. In a smaller subset (subgroup) of RA patients, anti-PAD4 antibodies can be measured and correlated with highly erosive forms of the disease.

  PAD4 inhibitors may also be useful in reducing neutrophil activity in pathologies in various diseases. Research has shown that neutrophil extracellular trap (NET) formation, the process of innate defense mechanisms by which neutrophils are immobilized and kill pathogens, is related to histone citrullination. And is suggested to be defective in PAD4 knockout mice (Neeli I. et. Al., J. Immunol., 180, (2008), 1895-1902 and Li P. et. Al., J. Exp. Med., 207 (9), (2010), 1853-1862). Thus, PAD4 inhibitors may have applicability to diseases where NET formation in tissues is involved in local injury and disease pathology. Such diseases include small vessel vasculitis (Kessenbrock K. et.al., Nat. Med., 15 (6), (2009), 623-625), systemic lupus erythematosus (Hakkim A. et.al. , Proc. Natl. Acad. Sci. USA, 107 (21), (2010), 9813-9818 and Villanuevar E. et. Al., J. Immunol., 187 (1), (2011), 538-52) Ulcerative colitis (Savchenko A. et. Al., Pathol. Int., 61 (5), (2011), 290-7), cystic fibrosis, asthma (Dworski R. et. Al., J. Allergy Clin. Immunol., 127 (5), (2011), 1260-6), deep vein thrombosis (Fuchs T. et. Al., Proc. Natl. Acad. Sci. USA, 107 (36), (2010 ), 15880-5), periodontal disease (Vitkov L. et. Al., Ultrastructural Pathol., 34 (1), (2010), 25-30), sepsis (Clark SR et al., Nat. Med., 13 ( 4), (2007), 463-9), appendicitis (Brinkmann V. et al., Science, 303, (2004), 1532-5) and stroke, but are not limited thereto. Further, NET is associated with diseases that affect the skin, such as the pathogenesis of cutaneous lupus erythematosus (Villanueva E. et al., J. Immunol., 187 (1), (2011), 538-52) and the pathogenesis of psoriasis (Lin AM et al., J. Immunol., 187 (1), (2011), 490-500), and therefore PAD4 inhibitors have been shown to address NET skin diseases when administered by systemic or dermal routes. Can show advantages. PAD4 inhibitors can affect further function in neutrophils and can have broader applicability to neutrophil related diseases.

  Studies have shown that collagen-induced arthritis (Willis VC et al., J. Immunol., 186 (7), (2011), 4396-4404), dextran sulfate sodium (DSS) -induced experimental colitis (Chumanevich A. A. et al., Am. J. Pysiol. Gastrointest. Liver Physiol., 300 (6), (2011), G929-G938), spinal cord repair (Lange S. et al., Dev. Biol., 355 (2), (2011 ), 205-14) and PAD inhibitors (eg, chloro-amidine) as a tool in several animal model diseases, including experimental autoimmune encephalomyelitis (EAE) ) Has been proven effective. DSS colitis reports also demonstrate that chloroamidine promotes apoptosis of inflammatory cells both in vitro and in vivo, and PAD4 inhibitors are more commonly used in a wide range of inflammatory diseases. It suggests that it can be effective.

  PAD4 inhibitors may also be useful in the treatment of cancer (Slack. J.L. et al., Cell. Mol. Life Sci., 68 (4), (2011), 709-720). Overexpression of PAD4 has been demonstrated in a number of cancers (Chang X. et al., BMC Cancer, 9, (2009), 40). The observation that PAD4 citrullinates arginine residues in histones at the promoter site of p53 target genes such as p21 involved in cell division cycle arrest and apoptosis induction suggests an antiproliferative role for PAD4 inhibitors It has been suggested (Li P. et al., Mol. Cell Biol., 28 (15), (2008), 4745-4758).

  The aforementioned role of PAD4 in deiminating arginine residues in histones can be an indicator of the role of PAD4 in regulating epigenetic gene expression. PAD4 is a primary PAD family member that is observed to be present in the nucleus as well as in the protoplasm. Early evidence that PAD4 can act as a histone demethyliminase as well as a deiminase is inconsistent and unproven. However, PAD4 may indirectly reduce histone arginine methylation by reducing the arginine residues that can be used by conversion to citrulline (ie, epigenetically regulated in relation to this goal). Do). Thus, PAD4 inhibitors may be useful as epigenetic tools or therapies that affect the expression of various target genes in additional disease situations. By such a mechanism, PAD4 inhibitors can also be effective in regulating the level of citrullination in stem cells, and thus a variety of, including but not limited to embryonic stem cells, neural stem cells, hematopoietic stem cells and cancer stem cells. Can have a therapeutic effect on the pluripotent constitution and differentiation potential of normal stem cells. Accordingly, there is an unmet need to identify and develop PAD4 inhibitors for the treatment of PAD4-mediated diseases.

(Summary of the Invention)
Compounds of formula I:

(Wherein ^{each of} ring A, R ¹ , R ² , R ³ , R ⁴ , X ¹ and X ² is as defined herein)
Alternatively, the pharmaceutically acceptable salt was found to be useful as an inhibitor of PAD4.

Compound of formula I ′:

Wherein each of ring A, ring B, R ¹ and R ³ is as defined herein.
Alternatively, it has been found that the pharmaceutically acceptable salt is useful as an inhibitor of PAD4.

  In certain embodiments, the present compounds exhibit selectivity for PAD4 over PAD2. The present invention also provides a pharmaceutically acceptable composition comprising the compound of the present application. The compounds of the present application are useful for the treatment of various diseases associated with PAD4. Such diseases are detailed herein, for example rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus and psoriasis.

(Detailed description of the invention)
1. General Description of Some Aspects of the Invention In certain embodiments, the compound is a formulation described herein, or a pharmaceutically acceptable salt thereof, wherein each variable in the formula Are as defined herein and in the embodiments. The compounds of the present invention have the formula I:
[Where
Ring A is

Wherein Ring A is 1-4 selected from fluorine, —CN, —OR or C _1-6 aliphatic groups (optionally substituted with 1 to 3 fluorine atoms). Optionally substituted with one group;
R ¹ is hydrogen, —CN, —OR or C _1-6 aliphatic group (optionally substituted with 1 to 4 groups selected from fluorine, —CN or OR);
R ² is hydrogen or a C _1-10 aliphatic group optionally substituted with 1 to 5 groups selected from fluorine, —CN or —OR;
Each of X ¹ and X ² is independently selected from N or C (R ⁴ );
Each of R ³ and R ⁴ is independently halogen, —CN, —R or —OR;
n is 0-4; and each R is independently hydrogen or a C _1-6 aliphatic group optionally substituted with 1 to 3 fluorine atoms.
Or a pharmaceutically acceptable salt thereof.

In certain embodiments, such compounds include compounds of the formulas described herein, or a pharmaceutically acceptable salt thereof, wherein each variable in the formula is defined herein, and Described in the embodiment. Such compounds have the formula I ′:
[Where
Ring A is



Wherein Ring A is _1-4 groups selected from fluorine, —CN, —OR or C _1-6 aliphatic groups (optionally substituted with 1 to 3 fluorine atoms) Optionally substituted with;
Ring B is

Is;
R ¹ is hydrogen, —CN, —OR,

Or a C _1-6 aliphatic group (optionally substituted with 1 to 4 groups selected from fluorine, —CN or —OR);
R ² is hydrogen or a C _1-10 aliphatic group optionally substituted with 1 to 5 groups selected from fluorine, —CN or —OR;
Each of X ¹ and X ² is independently selected from N or C (R ⁴ );
R ³ is halogen, —CN, —R,

Or -OR;
Each R ⁴ is independently halogen, —CN, —R,

Or -OR;
R ⁵ is hydrogen or halogen;
n is 0-4; and each R is independently hydrogen or a C _1-6 aliphatic group optionally substituted with 1 to 3 fluorine atoms.
Or a pharmaceutically acceptable salt thereof.

2. Definitions Compounds of the invention include those outlined herein and are further illustrated by the classifications, subclasses and types described herein. Unless otherwise specified, the following definitions as used herein apply. For the present invention, the chemical elements are, Handbook of Chemistry and Physics, 75 th periodic table of Ed, is specified according to CAS version. Additionally, general principles of organic chemistry, "Organic Chemistry", Thomas Sorrell , University Science Books, Sausalito:. 1999, and "March's Advanced Organic Chemistry", 5 th Ed, Ed .: Smith, MB and March, J. , John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

The term “aliphatic” or “aliphatic group” as used herein refers to a straight chain (ie, unbranched) or branched chain containing fully saturated or one or more unsaturated units. Means a branched or unsubstituted hydrocarbon chain, or contains fully saturated or one or more unsaturated units (but not aromatic) (herein “carbocycle”, Means a monocyclic or bicyclic hydrocarbon (also referred to as “alicyclic” or “cycloalkyl”) and having one point of attachment to the remainder of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In certain embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In another embodiment, aliphatic groups contain 1-4 aliphatic carbon atoms. In yet another embodiment, the aliphatic group contains 1 to 3 aliphatic carbon atoms, and in yet further embodiments, the aliphatic group contains 1 to 2 aliphatic carbon atoms. . In certain embodiments, “alicyclic” (or “carbocycle” or “cycloalkyl”) refers to a monocyclic C ₃ -C ₆ hydrocarbon that is fully saturated or attached to the remainder of the molecule. Refers to monocyclic C ₃ -C ₆ hydrocarbons containing one or more non-aromatic units having one point of attachment. Suitable aliphatic groups include straight or branched chain substituted or unsubstituted alkyl, alkenyl, alkynyl groups and mixtures thereof such as (cycloalkyl) alkyl, (cycloalkenyl) alkyl or (cycloalkyl) alkenyl. However, it is not limited to these.

  As used herein, the term “pharmaceutically acceptable salt” refers to human and lower animal tissues without undue toxicity, inflammation, allergic reactions, etc., within the scope of accurate medical judgment. It is suitable for use in contact with salt, and it meets a reasonable risk / benefit ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. Detail pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 1977, 66, 1-19, cited herein. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid or perchloric acid or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid. There are salts of amino groups formed with acids, succinic acid or malonic acid, or other methods used in the art, such as ion exchange methods. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate , Camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate Salt, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonic acid Salt, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, Acid salt, pectate salt, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p- Toluene sulfonate, undecanoate, valerate and the like are included.

Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include non-toxic ammonium, where appropriate formed with counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates. , Quaternary ammonium and amine cations.

Unless otherwise stated, structures shown herein are intended to cover all isomeric (eg, enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; It is also meant to include R and S configurations about the center, Z and E double bond isomers, and Z and E conformational isomers. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Further, unless otherwise stated, structures presented herein are also meant to include compounds that differ only in the presence of an atom enriched in one or more isotopes. For example, compounds having the structure of the present invention, including those in which hydrogen is replaced with deuterium or tritium, or carbons are replaced with ¹³ C- or ¹⁴ C-enriched carbon, Is within the range. The compounds are useful, for example, as analytical tools, probes in biological assays, or therapeutic agents according to the present invention.

  As used herein, the terms “measurably affinity” and “measurably inhibition” refer to PAD4, a compound of the invention or a composition thereof. As well as a measurable change in PAD4 activity between equivalent samples containing PAD4 in the absence of the compound or composition thereof.

3. DESCRIPTION OF EXEMPLARY COMPOUNDS According to one embodiment, the present invention provides compounds of formula I:
[Where
Ring A is

Wherein Ring A is _1-4 groups selected from fluorine, —CN, —OR or C _1-6 aliphatic groups (optionally substituted with 1 to 3 fluorine atoms) Optionally substituted with;
R ¹ is hydrogen, —CN, —OR or C _1-6 aliphatic group (optionally substituted with 1 to 4 groups selected from fluorine, —CN or —OR);
R ² is hydrogen or a C _1-10 aliphatic group (optionally substituted with 1 to 5 groups selected from fluorine, —CN or —OR);
Each of X ¹ and X ² is independently selected from N or C (R ⁴ );
Each of R ³ and R ⁴ is independently halogen, —CN, —R or —OR;
n is 0-4; and each R is independently hydrogen or a C _1-6 aliphatic group (optionally substituted with 1 to 3 fluorine atoms). ]
Or a pharmaceutically acceptable salt thereof.

According to another embodiment, the present invention provides compounds of formula I ′:
[Wherein ring A is



And ring A is 1-4 groups selected from fluorine, —CN, —OR or C _1-6 aliphatic groups (optionally substituted with 1 to 3 fluorine atoms). Optionally substituted;
Ring B is

Is;
R ¹ is hydrogen, —CN, —OR,

Or a C _1-6 aliphatic group (optionally substituted with 1 to 4 groups selected from fluorine, —CN or —OR);
R ² is a C _1-10 aliphatic group optionally substituted with 1 to 5 groups selected from fluorine, —CN or —OR;
Each of X ¹ and X ² is independently selected from N or C (R ⁴ );
R ³ is halogen, —CN, —R

Or -OR;
Each R ⁴ is independently halogen, —CN, —R,

Or -OR;
R ⁵ is hydrogen or halogen;
n is 0-4; and each R is independently hydrogen or a C _1-6 aliphatic group (optionally substituted with 1 to 3 fluorine atoms)]
Or a pharmaceutically acceptable salt thereof.

As defined above and as described herein, R ¹ is hydrogen, —CN, —OR or a C _1-6 aliphatic group (1-4 selected from fluorine, —CN or —OR Optionally substituted with a group). In certain embodiments, R ¹ is hydrogen. In certain embodiments, R ¹ is —CN. In certain embodiments, R ¹ is —OR. In certain embodiments, R ¹ is a C _1-6 aliphatic group optionally substituted with 1-4 groups selected from fluorine, —CN, or OR. In certain embodiments, R ¹ is a C _1-3 aliphatic group. In certain embodiments, R ¹ is methyl. In certain embodiments, R ¹ is ethyl. In certain embodiments, R ¹ is propyl. In certain embodiments, R ¹ is —CH ₂ -cyclobutyl, optionally substituted with methyl and —OH. In certain embodiments, R ¹ is
It is. In certain embodiments, R ¹ is

It is. In certain embodiments, R ¹ is

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It is. In certain embodiments, R ¹ is

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It is.

In certain embodiments, R ¹ is

It is. In certain embodiments, R ¹ is

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In certain embodiments, R ¹ is

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It is. In certain embodiments, R ¹ is

It is. In certain embodiments, R ¹ is

It is. In certain embodiments, R ¹ is selected from those listed in Table 1 below.

As defined above and as described herein, R ² is optionally substituted with 1-5 groups selected from hydrogen or C _1-10 aliphatic groups (fluorine, —CN or —OR). May be). In certain embodiments, R ² is hydrogen. In certain embodiments, R ² is a C _1-10 aliphatic group optionally substituted with 1 to 5 groups selected from fluorine, —CN or —OR. In certain embodiments, R ² is methyl. In certain embodiments, R ² is ethyl. In certain embodiments, R ² is propyl. In certain embodiments, R ² is butyl. In certain embodiments, R ² is pentyl. In certain embodiments, R ² is hexyl. In certain embodiments, R ² is cyclopropyl. In certain embodiments, R ² is cyclobutyl. In certain embodiments, R ² is cyclopentyl. In certain embodiments, R ² is cyclohexyl. In certain embodiments, R ² is cyclopropylmethyl. In certain embodiments, R ² is cyclobutylmethyl. In certain embodiments, R ² is cyclopentylmethyl. In certain embodiments, R ² is cyclohexylmethyl. In certain embodiments, R ² is cyclopropylethyl. In certain embodiments, R ² is cyclobutylethyl. In certain embodiments, R ² is cyclopentylethyl. In certain embodiments, R ² is cyclohexylethyl. In certain embodiments, R ² is —CH ₂ -cyclopropyl or —CH ₂ -cyclobutyl. In certain embodiments, R ¹ is —CH ₂ -cyclobutyl, optionally substituted with methyl and —OH.

In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 1 fluorine atom. In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 2 fluorine atoms. In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 3 fluorine atoms. In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 4 fluorine atoms. In certain embodiments, R ² is a C _1-10 aliphatic group substituted with 5 fluorine atoms. In certain embodiments, R ² is methyl substituted with 1 to 3 fluorine atoms. In certain embodiments, R ² is trifluoromethyl. In certain embodiments, R ² is ethyl substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is 2,2,2-trifluoroethyl. In certain embodiments, R ² is propyl substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is 3,3,3-trifluoropropyl. In certain embodiments, R ² is butyl substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is 4,4,4-trifluorobutyl. In certain embodiments, R ² is pentyl substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is 5,5,5-trifluoropentyl. In certain embodiments, R ² is hexyl substituted with 1 to 5 fluorine atoms. In certain embodiments, R ² is 6,6,6-trifluorohexyl. In certain embodiments, R ² is selected from those listed in Table 1 below.

As defined above and as described herein, each X ¹ and X ² is independently selected from N or C (R ⁴ ). In certain embodiments, both X ¹ and X ² are N. In certain embodiments, X ¹ is N and X ² is CH. In certain embodiments, X ¹ is CH and X ² is N. In certain embodiments, both X ¹ and X ² are CH. In certain embodiments, X ¹ is N and X ² is C (R ⁴ ). In certain embodiments, X ¹ is C (R ⁴ ) and X ² is N. In certain embodiments, both X ¹ and X ² are C (R ⁴ ). In certain embodiments, X ¹ and X ² are selected from those listed in Table 1 below.

As defined above and as described herein, R ³ is halogen, —CN, —R or —OR, each R independently being hydrogen or 1 to 3 fluorine atoms. An optionally substituted C _1-6 aliphatic group.

In certain embodiments, R ³ is hydrogen. In certain embodiments, R ³ is halogen. In certain embodiments, R ³ is —CN. In certain embodiments, R ³ is a C _1-3 aliphatic group. In certain embodiments, R ³ is methyl. In certain embodiments, R ³ is ethyl. In certain embodiments, R ³ is propyl. In certain embodiments, R ³ is —OR. In certain embodiments, R ³ is —OCH ₃ . In certain embodiments, R ³ is —OCH ₂ CH ₃ . In certain embodiments, R ³ is —OCH ₂ CH ₂ CH ₃ . In certain embodiments, R ³ is —OCH (F) ₂ .

In certain embodiments, R ³ is

It is. In certain embodiments, R ³ is

It is. In certain embodiments, R ³ is

It is. In certain embodiments, R ³ is selected from those listed in Table 1 below.

As defined above and as described herein, each R ⁴ is independently halogen, —CN, —R,

Or -OR.

In certain embodiments, R ⁴ is hydrogen. In certain embodiments, R ⁴ is halogen. In certain embodiments, R ⁴ is —CN. In certain embodiments, R ⁴ is C _1-6 aliphatic group or —OR. In certain embodiments, R ⁴ is —OCH ₃ . In certain embodiments, R ⁴ is ethyl.

In certain embodiments, R ⁴ is

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It is. In certain embodiments, R ⁴ is

It is. In certain embodiments, R ⁴ is selected from those listed in Table 1 below.

As defined above, ring A is

Wherein ring A is optionally substituted with 1 to 4 groups selected from fluorine or a C _1-6 aliphatic group optionally substituted with 1 to 3 fluorine atoms. Also good.

In certain embodiments, ring A is

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It is. In certain embodiments, Ring A is selected from those listed in Table 1 below.

As defined above and as described herein, ring B is

It is. In certain embodiments, ring B is

It is. In certain embodiments, ring B is

It is. In certain embodiments, ring B is

It is. In certain embodiments, ring B is

It is. In certain embodiments, Ring B is selected from those listed in Table 1 below.

As defined above and as described herein, R ⁵ is hydrogen or halogen. In certain embodiments, R ⁵ is hydrogen. In certain embodiments, R ⁵ is halogen. In certain embodiments, R ⁵ is fluoro. In certain embodiments, R ⁵ is selected from those listed in Table 1 below.

  As defined above and as described herein, n is 0-4. In certain embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In certain embodiments, n is selected from those listed in Table 1 below.

In certain embodiments, R ¹ is methyl, R ² is ethyl, X ¹ is N, X ² is CH, R ³ is hydrogen, and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is 2,2,2-trifluoroethyl, X ¹ is N, X ² is CH, and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is N, R ³ is —OCH ₃ , and ring A is

It is. In some embodiments, R ¹ is methyl, R ² is ethyl, X ¹ is N, X ² is CH, R ³ is hydrogen, and ring A is

is there. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is N, R ³ is hydrogen, and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is 2,2,2-trifluoroethyl, X ¹ is N, X ² is CH, and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is N, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is 2,2,2-trifluoroethyl, X ¹ is N, X ² is CH, and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is N, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, and ring A is

It is. In some embodiments, R ¹ is methyl, R ² is ethyl, X ¹ is N, X ² is CH, R ³ is hydrogen, and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, R ³ is —OCH ₃ , and ring A is
It is. In certain embodiments, R ¹ is methyl, R ² is cyclopropylmethyl, X ¹ is N, X ² is CH, R ³ is —OCH ₃ , and ring A is

It is. In certain embodiments, ring A is

It is.

  In certain embodiments, the compound of formula I is selected from those listed in Table 1.

Table 1. Exemplary compounds of formula I

  In certain embodiments, the present invention provides any compound described above or herein or a pharmaceutically acceptable salt thereof. In certain embodiments, the present invention provides a compound described in Table 1 above or a pharmaceutically acceptable salt thereof.

  In certain embodiments, the present invention provides any of the compounds described above and herein in isolated form.

4). According to another embodiment of the use, formulation and administration and pharmaceutically acceptable composition , the present invention comprises a compound of the invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant or Compositions comprising a vehicle are provided. The amount of the compound in the composition of the invention is an amount effective to inhibit PAD4 to a measurable extent in a biological sample or patient. In certain embodiments, the compositions of the invention are formulated for administration to a patient in need of the composition. In certain embodiments, the compositions of the invention are formulated for oral administration to a patient.

  As used herein, the term “subject” is used interchangeably with the term “patient” and means an animal, preferably a mammal. In certain embodiments, the subject or patient is a human. In another embodiment, the subject (or patient) is a veterinary subject (or patient). In certain embodiments, the veterinary subject (or patient) is a dog, cat or equivalent subject.

  The term “pharmaceutically acceptable carrier, adjuvant, or vehicle” refers to a non-toxic carrier, adjuvant, or vehicle that does not lose the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions of the invention include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as Phosphate, glycine, sorbic acid, potassium sorbate, partial glyceride mixture of saturated vegetable fatty acids, water, salt or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloid Silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic material, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, wax, polyethylene-polyoxypropylene-block polymer, polyethylene glycol and wool fat And the like.

  The compositions of the invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. As used herein, the term “parenteral” includes subcutaneous, intravenous, intramuscular, intraarticular, intrathecal, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. It is. Preferably, the composition is administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally used as a solvent or suspending medium.

  For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially polyoxyethylated ones thereof. is there. These oily solutions or suspensions may also contain similar dispersions commonly used in preparing pharmaceutically acceptable dosage forms, including long chain alcohol diluents or dispersants such as carboxymethylcellulose or emulsions and suspensions. An agent may be included. Other commonly used surfactants such as Tweens, Spans and other emulsifiers or bioavailability enhancers commonly used in making pharmaceutically acceptable solid, liquid or other dosage forms are also for formulation purposes. It may be used.

  The pharmaceutically acceptable compositions of this invention may be administered orally in any orally acceptable dosage form including capsules, tablets, aqueous suspensions or solutions. In the case of oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also usually added. Diluents useful for oral administration in a capsule form include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring, or coloring agents may also be added.

  Alternatively, the pharmaceutically acceptable compositions of this invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at the rectal temperature and therefore dissolves in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

  The pharmaceutically acceptable compositions of the present invention may be administered topically, especially when the target of treatment includes areas or organs that are readily reachable by topical use such as diseases of the eye, skin or lower intestinal tract. it can. Appropriate topical formulations are readily prepared for each of these areas or organs.

  Topical application of the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topical transdermal patches may be used.

  For topical application, provided pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for local administration of the compounds of the invention include mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions may be formulated in a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. . Suitable carriers include mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

  For ophthalmic applications, provided pharmaceutically acceptable compositions are fine powders in isotonic pH-adjusted sterile saline with or without preservatives such as benzylalkonium chloride. It may be formulated as a suspension or preferably as a solution in isotonic pH-adjusted sterile saline. Alternatively, for ophthalmic use, a pharmaceutically acceptable composition may be formulated in an ointment such as petrolatum.

  The pharmaceutically acceptable compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the pharmaceutical formulation art and are prepared in physiological saline with benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons and / or other conventional It can be prepared as a solution using a solubilizer or a dispersant.

  Most preferably, the pharmaceutically acceptable compositions of this invention are formulated for oral administration. Such formulations can be administered with or without a meal. In certain embodiments, the pharmaceutically acceptable compositions of this invention may be administered without a meal. In other embodiments, the pharmaceutically acceptable compositions of this invention are administered with meals.

  The pharmaceutically acceptable compositions of the present invention can be oral, rectal, parenteral, intracapsular, vaginal, intraperitoneal, topical (powder, ointment or infusion depending on the severity of the infection being treated. As an agent), it can be administered to humans and other animals, such as by mouth, oral or nasal spray. In certain embodiments, the compound of the present invention is from about 0.01 mg / kg to about 50 mg / kg, preferably from about 1 mg / kg to about 25 mg / kg per subject weight per day to achieve the desired therapeutic effect. It may be administered orally or parenterally once or multiple times daily at dosage levels.

  Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art such as water or other solvents, solubilizers and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl Alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene It may contain glycolic and sorbitan fatty acid esters and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants such as wetting, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

  Injectable preparations, for example, aqueous or oily sterile injection suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally used as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

  Injectable preparations are introduced with a sterilant, for example, by filtration through a bacteria-retaining filter or in the form of a sterile solid composition which can be dissolved or dispersed in sterile water or a sterile injectable medium before use. Can be sterilized.

  In order to sustain the effect of the compounds of the present invention, it is frequently desirable to delay the absorption of the compound by subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with low solubility in water. The absorption rate of a compound depends on its dissolution rate, which can depend on crystal size and crystal morphology. Alternatively, delaying the absorption rate of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the characteristics of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with living tissues.

  Compositions for rectal or vaginal administration can preferably be prepared by mixing a compound of the invention with a suitable nonirritating excipient or carrier such as cocoa butter, polyethylene glycols or suppository waxes. A suppository, which is solid at room temperature but liquid at body temperature, it dissolves in the rectum or vaginal cavity to release the active compound.

  Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In said solid dosage form, the active compound comprises at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or a) a filler or bulking agent such as Starch, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose and acacia, c) humectants such as glycerol, d) disintegrants such as agar, Calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) dissolution retardants such as paraffin, f) absorption enhancers such as quaternary ammonium compounds, g) wetting agents such as cetyl Alcohol and monostearies Glycerol acid, h) adsorbents such as kaolin and bentonite clays, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also contain buffering agents.

  Homogeneous solid compositions can be used as fillers for soft and hard gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycols. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifiers and only release the active ingredient (s) or preferentially release the active ingredient in a delayed manner, if desired, at specific sites in the intestine It can be present as a composition. Examples of embedding compositions that can be used include polymeric substances and waxes. Homogeneous solid compositions can also use excipients such as lactose or lactose and high molecular weight polyethylene glycols as fillers for soft and hard gelatin capsules.

  The active compound can take microencapsulated form with one or more excipients as described above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, controlled release coatings and other coatings well known in the pharmaceutical formulating art. In the solid dosage form, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. The dosage form may contain additional substances other than inert diluents, such as tableting lubricants and other tableting aids such as magnesium stearate and crystalline cellulose, as is usual practice. Good. In the case of capsules, tablets and pills, the dosage forms may also contain buffering agents. They may optionally contain opacifiers and only release the active ingredient (s) or preferentially release the active ingredient in a specific manner in the intestinal tract, optionally in a delayed manner. Can exist as a composition. Examples of embedding compositions that can be used include polymeric substances and waxes.

  Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers. Ophthalmic formulations, ear drops, and eye drops are also considered to be within the scope of this invention. Furthermore, the present invention contemplates the use of transdermal patches that have the additional advantage of controlled delivery of the compound to the body. The dosage form can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

  The amount of a compound of the present invention that can be combined with a carrier material to administer the composition in a single dosage form will vary depending upon the host treated and the particular mode of administration. Preferably, the provided composition should be formulated such that a dosage of 0.01-100 mg / kg body weight / day can be administered to a patient taking the composition.

  The compounds of the invention can be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapies take the form of fixed combinations or, independently of each other, One or more therapeutic compounds with a fixed combination or a fixed combination, optionally in the form of administration of a compound of the present invention and one or more other therapeutic compounds at a given time The administration form is combined. In addition or in addition, the compounds of the invention can be administered in particular for chemotherapy, in particular chemotherapy, radiation therapy, immunotherapy, phototherapy, surgical treatment, or a combination thereof. Long-term therapy is equally possible as an adjunct therapy in the context of other treatment modalities, as described above. Other possible treatments are treatments to maintain the patient's condition after tumor regression or, for example, after prophylactic chemotherapy in a patient at risk.

  These additional agents may be administered separately from the compound-containing composition of the invention as part of multiple dosing regimens. Alternatively, these agents can be part of a single dosage form that is mixed with a compound of the invention in a single composition. When administered as part of a multiple dosing regimen, the two active agents may be provided simultaneously, sequentially or within a fixed time of each other, usually within 5 hours of each other.

  As used herein, the terms “combination”, “combined” and related terms refer to simultaneous or sequential administration of the therapeutic agents described in the present invention. For example, the compounds of the present invention can be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present invention provides a single unit dosage form comprising a compound of the present invention, an additional therapeutic agent and a pharmaceutically acceptable carrier, adjuvant or vehicle.

  The compound of the invention and additional therapeutic agent (in its composition comprising the additional therapeutic agent described above) combined with a carrier material to administer a single dosage form will vary depending on the host being treated and the particular mode of administration. Will do. Preferably, the compositions of the invention should be formulated so that a dosage of 0.01-100 mg / kg body weight / day of the compound of the invention can be administered.

  In compositions comprising an additional therapeutic agent, the additional therapeutic agent and the compound of the invention can act synergistically. Thus, the amount of additional therapeutic agent in the composition is less than that required for monotherapy using only that therapeutic agent.

  The amount of additional therapeutic agent present in the composition of the present invention will be no more than the amount normally administered in a composition comprising that therapeutic agent as the only active agent. Preferably, the amount of additional therapeutic agent in the composition according to the invention ranges from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent. .

  The particular dosage and treatment regimen for any particular patient will depend on various factors such as the activity of the particular compound used, age, weight, general health, sex and diet, administration time, elimination rate, drug It will be appreciated that this will depend on the combination of and the judgment of the attending physician and the severity of the particular disease being treated. The amount of the compound of the invention in the composition will also depend on the particular compound in the composition.

Compound Use and Pharmaceutically Acceptable Compositions The compounds and compositions described herein are generally useful for the inhibition of PAD4.

  The activity of the compounds used in the present invention as inhibition of PAD4 can be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that measure inhibition of PAD4. Detailed conditions for assaying the compounds used in the present invention as inhibitors of PAD4 are as described in the Examples below. In certain embodiments, the compounds of the present invention selectively inhibit PAD4 relative to PAD2.

  As used herein, the terms “treatment”, “treating” and “treating” refer to whether the disease or disorder described herein or one or more symptoms thereof are ameliorated or alleviated. , Delaying its onset or inhibiting its progression. In certain embodiments, treatment may be administered after onset of one or more symptoms. In other embodiments, treatment may be given when there are no symptoms. For example, treatment may be given to patients who are susceptible to infection prior to the onset of symptoms (eg, taking into account the history of symptoms and / or genetic or other susceptibility factors). Treatment may continue after symptoms disappear, for example, to prevent or delay their recurrence.

  The compounds of the present invention are inhibitors of PAD4 and are therefore useful for treating one or more diseases associated with the activity of PAD4. Accordingly, in one embodiment, the invention provides a method of treating a PAD4-mediated disease, comprising the step of administering a compound of the invention or a pharmaceutically acceptable composition thereof to a patient in need thereof. provide.

  In certain embodiments, the PAD4-mediated disease is a disease, symptom or disorder mediated by inappropriate PAD4 activity. In certain embodiments, the PAD4-mediated disease is selected from the group consisting of rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus and psoriasis. In a further embodiment, the disease mediated by inappropriate PAD4 activity is rheumatoid arthritis. In a further embodiment, the disease mediated by inappropriate PAD4 activity is systemic lupus erythematosus. In a further embodiment, the disease mediated by inappropriate PAD4 activity is vasculitis. In a further embodiment, the disease mediated by inappropriate PAD4 activity is cutaneous lupus erythematosus. In a further embodiment, the disease mediated by inappropriate PAD4 activity is psoriasis.

  In one embodiment, a method for treating rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus or psoriasis is provided, the method comprising: A therapeutically effective amount of the present compound or a pharmaceutically acceptable salt thereof is administered to a human subject with

  In one embodiment, a method of treating rheumatoid arthritis is provided, which comprises administering to a human subject in need thereof a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof. It is characterized by. In one embodiment, a method of treating systemic lupus erythematosus is provided, which method comprises administering to a human subject in need thereof a therapeutically effective amount of the present compound or a pharmaceutically acceptable salt thereof. It is characterized by that. In one embodiment, a method for treating vasculitis, comprising administering to a human subject in need thereof a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof. It is characterized by. In one embodiment, a method of treating cutaneous lupus erythematosus is provided, the method comprising administering to a human subject in need thereof a therapeutically effective amount of the present compound or a pharmaceutically acceptable salt thereof. It is characterized by. In one embodiment, a method for treating psoriasis comprising administering to a human subject in need thereof a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof. Features.

  In certain embodiments, the PAD4-mediated disease is acid-induced lung injury, acne (PAPA), acute lymphoblastic leukemia, acute respiratory distress syndrome, Addison's disease, adrenal hyperplasia, adrenal cortical dysfunction, aging, AIDS, alcohol Hepatitis, alcoholic hepatitis, alcoholic liver disease, allergic asthma, allergic bronchopulmonary aspergillosis, allergic conjunctivitis, alopecia, Alzheimer's disease, amyloidosis, amyotrophic lateral sclerosis and weight loss, angina, Angioedema, nonhidrotic ectodermal dysplasia (AHED-ID), ankylosing spondylitis, anterior segment, inflammation, antiphospholipid syndrome, aphthous stomatitis, appendicitis, arthritis, asthma, atheroma Atherosclerosis, atopic dermatitis, autoimmune disease, autoimmune hepatitis, bee sting-induced inflammation, Behcet's disease, Behcet's syndrome, bell paralysis, Lilium poisoning, Blau syndrome, bone pain, bronchiolitis, burns, bursitis, cancer, cardiac hypertrophy, carpal tunnel syndrome, cataract disorder, cataract, cerebral aneurysm, chemical stimulus-induced inflammation, choroidal retina Inflammation, chronic heart disease, chronic lung disease in premature infants, chronic lymphocytic leukemia, chronic obstructive pulmonary disease, colitis, complex local pain syndrome, collagen disease, corneal ulcer, Crohn's disease, cryopyrin-related periodic fever syndrome, cryptococcosis Cystic fibrosis, interleukin-1-receptor antagonist (DIRA) deficiency, dermatitis, dermatitis endotoxemia, dermatomyositis, childhood brain stem glioma, endometriosis, endotoxemia, epicondyle Inflammation, erythrocytopenia, familial amyloid polyneuropathy, familial cold urticaria, familial Mediterranean fever, fetal growth retardation, glaucoma, glomerular disease, glomerulonephritis, gout, gouty arthritis, graft pair Host disease, digestive system disease, head Traumatic injury, headache, hearing loss, heart disease, hemolytic anemia, Henoch-Schorain purpura, hepatitis, hereditary autoinflammatory syndrome, herpes zoster and herpes simplex, HIV-1, Hodgkin's disease, Huntington's disease, pulmonary hyaline, Hyperammonemia, hypercalcemia, hypercholesterolemia, hyperimmunoglobulinemia D syndrome with periodic fever (HIDS), aplastic and other anemias, aplastic anemia, idiopathic thrombocytopenic purpura Disease, dyschromia, infectious mononucleosis, inflammatory bowel disease, inflammatory lung disease, inflammatory neuropathy, inflammatory pain, insect bite-induced inflammation, iritis, irritation-induced inflammation, ischemia / Reperfusion, juvenile rheumatoid arthritis, keratitis, kidney disease, kidney damage caused by parasitic infection, kidney damage causing parasitic infection, prevention of kidney transplant rejection, leptospirosis, leukemia, Lehler syndrome, lung injury, lung injury , Lupus, wolf Pressure ulcer, lupus nephritis, lymphoma, meningitis, mesothelioma, mixed collagen disease, Maccle Wells syndrome (deafness amyloidosis of hives), multiple sclerosis, muscle fatigue, muscular dystrophy, myasthenia gravis, myocarditis, Mycosis fungoides, mycosis fungoides, myelodysplastic syndrome, myositis, rhinosinusitis, necrotizing enterocolitis, neonatal onset multiple organ inflammatory disease (NOMID), nephrotic syndrome, neuritis, neuropathology Disease, non-allergy-induced asthma, obesity, ocular allergy, optic neuritis, organ transplantation, osteoarthritis, otitis media, Paget's disease, pain, pancreatitis, Parkinson's disease, pemphigus, pericarditis, periodic fever, periodontitis Peritoneal endometriosis, pertussis, sore throat and lymphadenitis (PFAPA syndrome), plant stimulation-induced inflammation, pneumonia, pneumonitis, pneumonitis, pneumocystis pneumonia, American poison ivy / urushiol oil-induced inflammation, Nodularity Arterial, polychondritis, polycystic kidney disease, polymyositis, psoriasis, psoriasis, psoriasis, psoriasis, psychosocial stress disease, lung disease, pulmonary hypertension, pulmonary fibrosis, gangrenous pyoderma, purulent Arthritis, kidney disease, retinal disease, rheumatic carditis, rheumatic disease, rheumatoid arthritis, sarcoidosis, seborrhea, sepsis, severe pain, sickle cell, sickle cell anemia, silica-induced disease, Sjogren's syndrome, skin disease , Sleep apnea, solid tumor, spinal cord injury, Stevens-Johnson syndrome, stroke, subarachnoid hemorrhage, sunburn, temporal arteritis, tendon synovitis, thrombocytopenia, thyroiditis, tissue transplantation, TNF receptor From associated periodic syndrome (TRAPS), toxoplasmosis, transplantation, traumatic brain injury, tuberculosis, type I diabetes, type II diabetes, ulcerative colitis, urticaria-like uveitis, uveitis and Wegener's granulomatosis It is selected from that group.

  In one embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in therapy. In another embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of a disease mediated by inappropriate PAD4 activity. In another embodiment, the present invention provides a compound of the present invention or a medicament thereof for use in the treatment of rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus or psoriasis A chemically acceptable salt. In another embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of rheumatoid arthritis. In another embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of systemic lupus erythematosus. In another embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of vasculitis. In another embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of cutaneous lupus erythematosus. In another embodiment, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of psoriasis. In another embodiment, the present invention provides the use of the present compounds or pharmaceutically acceptable salts thereof in the manufacture of a medicament for use in the treatment of diseases mediated by inappropriate PAD4 activity. In another embodiment, the present invention relates to a compound of the invention in the manufacture of a medicament for use in the treatment of rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus or psoriasis. Alternatively, the use of a pharmaceutically acceptable salt thereof is provided. In another embodiment, the present invention provides the use of the present compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of rheumatoid arthritis. In another embodiment, the present invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of systemic lupus erythematosus. In another embodiment, the present invention provides the use of the present compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of vasculitis. In another embodiment, the present invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of cutaneous lupus erythematosus. In another embodiment, the present invention provides the use of the present compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of psoriasis. In a further embodiment, the present invention provides a pharmaceutical composition comprising the present compound or a pharmaceutically acceptable salt thereof for the treatment or prevention of a disease mediated by inappropriate PAD4 activity. In a further embodiment, the present invention provides a compound of the present invention or a medicament thereof for the treatment or prevention of rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus or psoriasis A pharmaceutical composition comprising a pharmaceutically acceptable salt is provided. In a further embodiment, the present invention provides a pharmaceutical composition comprising the present compound or a pharmaceutically acceptable salt thereof for the treatment or prevention of rheumatoid arthritis. In a further embodiment, the present invention provides a pharmaceutical composition comprising the present compound or a pharmaceutically acceptable salt thereof for the treatment or prevention of systemic lupus erythematosus. In a further embodiment, the present invention provides a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof for the treatment or prevention of vasculitis. In a further embodiment, the present invention provides a pharmaceutical composition comprising the present compound or a pharmaceutically acceptable salt thereof for the treatment or prevention of cutaneous lupus erythematosus. In a further embodiment, the present invention provides a pharmaceutical composition comprising the present compound or a pharmaceutically acceptable salt thereof for the treatment or prevention of psoriasis.

  All features of each aspect of the invention apply mutatis mutandis to all other aspects.

  In order that the invention described herein may be more fully understood, the following examples are set forth below. It should be understood that these examples are for illustrative purposes and are not intended to limit the invention in any manner.

Exemplification In the illustration of certain embodiments, as shown in the examples below, the compounds were prepared according to the following general method. The general methods represent the synthesis of certain compounds of the invention, but the following general methods and alternatives known to those skilled in the art include all compounds as described herein and each of these compounds. It will be understood that this can be applied to subclasses and strains.

Method A
MET / u-HPLC (low pH 7 min method)
Column: Phenomenex Kinetex-XB C18, 2.1mm x 100mm, 1.7μm
Flow rate: 0.6 ml / min Mobile phase:
A: 0.1% formic acid (aqueous solution) and B: 0.1% formic acid (MeCN)
Injection volume: 3 μl
Temperature: 40 ° C
Detection: 215 nm (nominal)
Gradient time (min)-% B
0.00-5
5.30-100
5.80-100
5.82-5

Method B
MET / CR / 1600 (High pH 7 minutes method)
Column: Phenomenex Gemini C18, 2.0mm x 100mm, 3μm
Flow rate: 0.5 ml / min Mobile phase:
A: 2 mM ammonium bicarbonate / HPLC grade water pH 10
B: HPLC grade MeCN
Injection volume: 3 μl
Temperature: 50 ° C
Detection: 215nm
Gradient time: (min)-% B
1.0-5
5.50-100
5.90-100
5.92-5
9.00-5

Method C
METCR 1416 (Low pH Shimadzu 7 min method)
Column: Waters Atlantis dC18, 2.1 mm x 100 mm, 3 μm Column flow rate: 0.6 ml / min Mobile phase:
A: 0.1% formic acid (aqueous solution) and B: 0.1% formic acid (acetonitrile)
Injection volume: 3 μl
Temperature: 40 ° C
Detection: 215 nm (nominal)
Gradient time (min)-% B
0.00-5
5.100 to 100
5.40-100
5.42-5

Method D
METCR 1410 (Low pH Shimadzu 2 minutes method)
Column: Kinete x Core-Shell C18, 2.1 mm x 50 mm, 5 μm Column flow rate: 1.2 ml / min Mobile phase:
A: 0.1% formic acid (aqueous solution) and B: 0.1% formic acid (acetonitrile)
Injection volume: 3 μl
Temperature: 40 ° C
Detection: 215 nm (nominal)
Gradient time (min)-% B
0.00-5
1.20-100
1.30-100
1.31-5

Method E:
Chiral HPLC preparative method Column: Chiralpak IC 250 mm x 4.6 mm, 5 μm Column flow rate: 15 ml / min Mobile phase: 35% Ethanol: 65% CO ₂
Sample diluent: ethanol temperature; 40 ° C
Detection: 215 nm (nominal)

Method F:
Kira purity analysis method Column: Chiralpak IC 250mm x 4.6mm, 5μm Column flow rate: 4ml / min Injection volume: 10μL
Temperature: 40 ° C
Detection: 215nm
Isocratic conditions 40% ethanol: 60% CO ₂

Method G
Chiral HPLC preparative column: XSelect CSH C18 50 x 2.1 mm, 1.7um
Flow rate: 0.6 ml / min Mobile phase: water (0.1% v / v TFA), MecN (0.1% v / v TFA)
Sample diluent: ethanol temperature; 40 ° C
Detection: 240nm (nominal)

Method H
MET / u-HPLC (High pH MS16 7 min method)
Column: Waters UPLC CSH C18, 2.1 mm x 100 mm 5 μm Column flow rate: 0.6 ml / min Mobile phase:
A: Ammonium hydrogen carbonate salt adjusted to 2 mM pH 10 (including ammonium hydroxide (aqueous solution)), B: Acetonitrile injection amount: 3 μl
Temperature: 40 ° C
Detection: 215 nm (nominal)
Gradient time (min)-% B
0.00-5
5.30-100
5.80-100
5.82-5

Method I:
Kira purity analysis method column: Lux C4 (21.2mm x 250mm, 5um)
Flow rate: 21 ml / min Injection amount: 350 μL
Detection: 222nm
Isocratic conditions: MeOH (0.1% v / vNH ₃ )

Method J
MET / CR / 0990 (High pH 3 minutes method)
Column: Phenomenex Gemini C18, 2.0mm x 100mm, 3μm
Flow rate: 1 ml / min Mobile phase:
A: 2 mM ammonium bicarbonate / HPLC grade water pH 10
B: HPLC grade MeCN
Injection volume: 3 μl
Temperature: 60 ° C
Detection: 215nm
Gradient time: (min)-% B
0.0-1
1.80-100
2.10-100
2.30-1

Method K:
Chiral HPLC preparative column: Amy-C 20mm x 250mm, 5um
Flow rate: 21 ml / min Mobile phase:
4: 6 Heptane: ethanol (0.1% v / v ammonia)
Sample diluent: methanol temperature; ambient temperature detection: 254 nm

Method L:
Kira Purity Analysis Method Column: Amy-C 4.6mm x 250mm, 5um
Flow rate: 21 ml / min Injection volume: 1.0 μL
Temperature; ambient temperature UV detection: 254 nm
Isocratic conditions, 4: 6 heptane: ethanol (0.1% v / v ammonia)

Certain compounds of the present invention were prepared according to Scheme 1 below.

2- {1-Ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6 -Carbonyl} -1H-1,3-benzodiazole EV-AR0076-002 (EOAI 3428370), Synthesis of I-13

Methyl 3-methoxy-4- (methylamino) -5-nitrobenzoate EV-AR0065-002-step 1
To a stirred solution of methyl 4-chloro-3-methoxy-5-nitrobenzoate (CAS 63603-09-8, 2.0 g, 8.14 mmol) / DMF (10 ml) was added K ₂ CO ₃ (99%, 1.37 g, 9.81 mmol) was added. To this solution was added methanamine hydrochloride (1: 1) (0.62 g, 9.18 mmol) and the mixture was stirred in a sealed tube at 80 ° C. for 16 hours under nitrogen. The reaction crude was concentrated in vacuo and partitioned between DCM (100 ml) and water (10 ml). The organic layer was further washed with water (2 × 10 ml) and saturated aqueous sodium chloride solution (10 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give an orange powder that was purified by flash chromatography (15-40% EtOAc / heptane) and methyl 3-methoxy. -4- (Methylamino) -5-nitrobenzoate EV-AR0065-002 [1.49 g (76%)] was obtained as an orange powder. LCMS (Method D): retention time 1.13 minutes, M / z = 241- (M + 1).

Methyl 3-amino-5-methoxy-4- (methylamino) benzoate EV-AR0068-002-step 2
To a stirred solution of methyl 3-methoxy-4- (methylamino) -5-nitrobenzoate (EV-AR0065-002, 1.49 g, 6.20 mmol) / ethanol (100 ml) under nitrogen was added 10% Pd / C (0.18 g, 0.17 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction mixture was filtered through Kieselguhr and the filtrate was washed through methanol (150 ml). The filtrate was concentrated in vacuo to give methyl 3-amino-5-methoxy-4- (methylamino) benzoate EV-AR0068-002 [1.21 g (89%)] as a pale purple powder. LCMS (Method D): retention time 0.63 minutes, M / z = 211− (M + 1).

Ethyl 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AQ1957-001-Step 3
Sodium hydride (60%, 59 mg, 1.47 mmol) was added dropwise in several portions and at room temperature ethyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate (CAS 221675-35- 0,200 mg, 1.05 mmol) / DMF (5 ml) was added. The mixture was stirred for 20 minutes and then ethyl iodide (197 mg, 1.26 mmol) was added. The reaction mixture was stirred for 20 hours. The mixture was partitioned between EtOAc (20 ml) and water (20 ml). The aqueous layer was further extracted with EtOAc (1 × 20 ml) and the combined organic layers were washed with water (20 ml) and evaporated to dryness. The crude product was purified by flash chromatography (0-50% EtOAc / heptane) to give ethyl 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AQ1957-001 [135 mg. (57.1%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.18 minutes, M / z = 219 (M + 1).

  1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AQ1960-001-step 4

  To a stirred solution of ethyl 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AQ1957-001, 135 mg, 0.62 mmol) / THF (2 ml) was added lithium hydroxide (74 mg, 3.09 mmol) / water (2 ml) was added. The mixture was stirred at 50 ° C. for 2.5 hours. The mixture was acidified with 1M HCl (3 ml) and extracted with DCM (2 × 5 ml). The combined organics were washed with water and evaporated to dryness to give 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AQ1960-001 [120 mg (99%)] as a white solid. Obtained. LCMS (Method D): retention time 0.92 min, M / z = 191- (M + 1).

Methyl 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AR0070 -003-Step 5
To a stirred solution of 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AQ1960-001, 120 mg, 0.63 mmol) / DMF (2 ml) was added DIPEA (116 μl, 0.70 mmol). ), Then HATU (236 mg, 0.62 mmol) was added and the resulting mixture was stirred at room temperature for 15 minutes. Methyl 3-amino-5-methoxy-4- (methylamino) benzoate (EV-AR0068-002, 148 mg, 0.70 mmol) was added and the resulting mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated in vacuo, dissolved in acetic acid (3 ml) and stirred at 70 ° C. for 16 hours. The solvent was removed in vacuo and purified by flash chromatography (25-40% EtOAc / heptane) to give methyl 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl}- 7-Methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AR0070-003 [150 mg (63%)] was obtained as a white powder. LCMS (Method D): retention time 1.18 minutes, M / z = 365 (M + 1).

2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid EV-AR0072- 002-Step 6
Methyl 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate (EV- To a stirred solution of AR0070-003, 150 mg, 0.41 mmol) / THF (3 ml) was added a solution of lithium hydroxide (30 mg, 1.25 mmol) / water (3 ml) and the mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated in vacuo, taken up in water (5 ml) and acidified with 5N HCl (0.5 ml) with stirring. The resulting suspension was stirred for 10 minutes, then the precipitate was collected by vacuum filtration and dried to give 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridine-2- Yle} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid EV-AR0072-002 [130 mg (89%)] was obtained as a white powder. LCMS (Method D): retention time 1.03 min, M / z = 351- (M + 1).

  Tert-butyl 6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 Carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0074-002-Step 7

  2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1, in 2: 1 DMSO / MeCN (4.5 ml), To a stirred solution of 3-benzodiazole-5-carboxylic acid (EV-AR0072-002, 130 mg, 0.37 mmol) was added DIPEA (65 μl, 0.39 mmol) and HATU (148 mg, 0.39 mmol). The resulting mixture was stirred at room temperature for 15 minutes, then tert-butyl octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (CAS 949559-11-9,88 mg, 0.39 mmol) was added. The mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with 3: 2 MeCN / water (1.5 ml) and purified by preparative HPLC (basic method) to give tert-butyl 6- (2- {1-ethyl-1H- Pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] Pyridine-1-carboxylate EV-AR0074-002 [142 mg (81%)] was obtained as a white powder. LCMS (Method A): retention time 3.41 min, M / z = 559 (M + 1).

  2- {1-Ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6 -Carbonyl} -1H-1,3-benzodiazole EV-AR0076-002 (EOAI 3428370), I-13-step 8

  tert-butyl 6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 To a stirred solution of carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AR0074-002, 20 mg, 0.04 mmol) / methanol (1 ml) 4M HCl / 1,4 -Dioxane (0.5 ml) was added and the resulting solution was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuo and the residue was lyophilized from water (4 ml) to give 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy. -1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl} -1H-1,3-benzodiazole hydrochloride EV-AR0076-002, I-13 [13. 4 mg (75%)] was obtained as a yellow solid. LCMS (Method A): retention time 1.80 minutes, M / z = 459 (M + 1).

  tert-Butyl (3aR, 7aS) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0090-001 and tert-butyl (3aS, 7aR) -6- (2- {1- Ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2, 3-c] Pyridine-1-carboxylate Chiral HPLC to obtain EV-AR0090-002—Step 9

tert-butyl 6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 Carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0074-002 [107 mg] was dissolved in ethanol and then purified by chiral HPLC (Method E) to give tert -Butyl (3aR, 7aS) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzo Diazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0090-001 (absolute stereochemistry assigned arbitrarily) [46.6 mg (43.6 %)] And tert-buty (3aS, 7aR) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodi Azole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0090-002 (absolute stereochemistry assigned arbitrarily) [36.8 mg (33.4% )].

EV-AR0090-001, chiral purity (UV, 254 nm): 100%, retention time: 6.30 minutes (Method F)

EV-AR0090-002, Chiral purity (UV, 254 nm): 97%, Retention time: 9.96 minutes (Method F)

5-[(3aS, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridin-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl } -7-Methoxy-1-methyl-1H-1,3-benzodiazole EV-AR0091-002 (EOAI 3342499, absolute stereochemistry assigned arbitrarily) I-3-Step 8
Tert-butyl (3aR, 7aS) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AR0090-001, 46 mg, 0.08 mmol) as in step 8, scheme 1 5-[(3aS, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b] pyridine -2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole hydrochloride EV-AR0091-002, I-3 [40 mg (97%)] was obtained as an orange powder. LCMS (Method A): retention time 1.83 minutes, M / z = 459 (M + 1).

  5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridin-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl } -7-Methoxy-1-methyl-1H-1,3-benzodiazole EV-AR0092-002 (EOAI 3432500, absolute stereochemistry assigned arbitrarily) I-14-Step 8

  Tert-butyl (3aS, 7aR) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AR0090-002, 36.8 mg, 0.07 mmol) was prepared according to step 8, scheme 1. 5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b ] Pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole EV-AR0092-002, I-14 [31 mg (92.7%)] was obtained as an orange powder. . LCMS (Method A): retention time 1.82 minutes, M / z = 459 (M + 1).

Scheme 2

1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl}- Synthesis of 4-methoxypiperidin-3-amine EV-AS1585-001- (EOAI3436357), I-62

1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AR3164-001-Step 1
To a stirred solution of ethyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate (CAS 221675-35-0, 4.40 g, 23.1 mmol) / DMF (50 ml) was added sodium hydride (60%, 1 .05 g, 26.3 mmol) was added. The mixture was stirred at room temperature for 45 minutes under nitrogen and (bromomethyl) cyclopropane (CAS7051-34-5, 2.70 ml, 27.8 mmol) was added. The mixture was stirred at room temperature for 2.5 hours and the solvent was removed in vacuo. The residue was suspended in THF (40 ml) and 5M aqueous sodium hydroxide (22 ml, 110 mmol) was added. The mixture was stirred at 50 ° C. for 3.5 hours. Additional THF (20 ml) and 5M aqueous sodium hydroxide (22 ml, 110 mmol) were added and the reaction was stirred at 50 ° C. for 16 hours. The reaction crude product was concentrated in vacuo and water (10 ml) and 5M aqueous hydrochloric acid (100 ml) were added. The solid was filtered off, washed with water (2 × 100 ml), dried in a vacuum oven and 1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AR3164- 001 [3.46 g (69.2%)] was obtained as a white powder. LCMS (Method D): retention time 1.03 min, M / z = 217 (M + 1).

Methyl 4- (methylamino) -3-nitrobenzoate EV-AR3152-001-Step 2
To a stirred solution of methyl 4-fluoro-3-nitrobenzoate (CAS329-59-9,5.00 g, 25.1 mmol) / DMF (50 ml) was added methanamine hydrochloride (1: 1) (2.00 g, 29.6 mmol). ) And potassium carbonate (4.50 g, 32.6 mmol). The mixture was stirred at room temperature for 18 hours under nitrogen. The crude product of the reaction was concentrated in vacuo and the residue was partitioned between EtOAc (350 ml) and 1N aqueous hydrochloric acid (250 ml). The organic layer was further washed with 1N aqueous hydrochloric acid (150 ml) and saturated aqueous sodium chloride (100 ml). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give methyl 4- (methylamino) -3-nitrobenzoate EV-AR3152-001 [5.30 g (quantitative)] as a yellow powder. Got as. LCMS (Method D): retention time 1.07 minutes, M / z = 211− (M + 1).

Methyl 3-amino-4- (methylamino) benzoate EV-AR3155-001-step 3
To a stirred solution of methyl 4- (methylamino) -3-nitrobenzoate (EV-AR3152-001, 5.30 g, 25.2 mmol) / ethanol (100 ml) was added 10% Pd / C (1.30 g under nitrogen). , 0.05 mmol). The reaction was then stirred at room temperature for 4 hours under a hydrogen atmosphere. The reaction mixture was diluted with methanol (100 ml) and Kieselguhr was added. The mixture was stirred at room temperature for 10 minutes and vacuum filtered. The filtrate was washed with methanol (3 × 50 ml) and the filtrate was concentrated in vacuo to give methyl 3-amino-4- (methylamino) benzoate EV-AR3155-001 [4.39 g (96.6%)] as a brown powder. It was. LCMS (Method D): retention time 0.75 min, M / z = 181- (M + 1).

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AR3167 -001-Step 4

  To a solution of 1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AR3164-001, 2.20 g, 10.2 mmol) / dry DMF (40 ml) was added HATU. (4.95 g, 12.8 mmol) and DIPEA (2.25 ml, 12.8 mmol) were added. The mixture was stirred at room temperature for 1 hour and then methyl 3-amino-4- (methylamino) benzoate (EV-AR3155-001, 2.02 g, 11.2 mmol) was added. The mixture was stirred at room temperature for 16 hours. The solvent was removed in vacuo and the residue was dissolved in acetic acid and stirred at 80 ° C. for 2 hours, then heated at 85 ° C. for 30 minutes and then at 90 ° C. for 1 hour. The solvent was removed in vacuo and the crude material was purified by flash chromatography (12-100% EtOAc / heptane) to give methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b. ] Pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AR 3167-001 [3.08 g (83.2%)] was obtained as a pink powder. It was. LCMS (Method D): retention time 1.20 minutes, M / z = 361- (M + 1).

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-5-carboxylic acid EV-AR3168- 002-Step 5

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-5-carboxylate (EV- To a suspension of AR3167-001, 3.08 g, 8.46 mmol) / methanol (60 ml) was added 2M aqueous sodium hydroxide solution (30 ml, 60.0 mmol). The mixture was then stirred at 50 ° C. for 2 hours. The reaction solution was cooled to room temperature and the solvent was removed in vacuo. Water (50 ml) was added followed by 2M aqueous HCl until pH 3 was achieved. The mixture was stirred for 15 minutes and filtered through sinter. The solid was filtered through water (2 × 50 ml) and air dried for 64 hours to give 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H. -1,3-Benzodiazole-5-carboxylic acid EV-AR3168-001 [1.81 g (61.2%)] was obtained as a flesh-colored solid. LCMS (Method D): retention time 1.05 min, M / z = 347 (M + 1). The filtrate was acidified by adding more 2M aqueous HCl until a precipitate began to form. The mixture was allowed to stand for 1 hour and filtered through sinter. The solid was washed with water (2 × 20 ml) and air dried under vacuum for 3 hours to give 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl- 1H-1,3-benzodiazole-5-carboxylic acid EV-AR3168-002 [460 mg, (15.7%)] was obtained as an off-white powder. LCMS (Method D): retention time 1.06 minutes, M / z = 347 (M + 1).

  Benzyl (3S, 4R) -3-{[(tert-butoxy) carbonyl] amino} -4-methoxypiperidine-1-carboxylate EV-AT1700-001-Step 6

  Benzyl (3S, 4R) -3-{[(tert-butoxy) carbonyl] amino} -4-hydroxypiperidine-1-carboxylate (EV-AQ 1997-001, 450 mg, 1.28 mmol) synthesized as WO 2014/015905 ) / Anhydrous THF (10 ml) was added sodium hydride (60%, 62 mg, 1.54 mmol) at 0 ° C. under nitrogen. The mixture was stirred at 0 ° C. for 30 minutes, then iodomethane (83.94 μl, 1.35 mmol) was added and the mixture was stirred at room temperature for 16 hours. Saturated aqueous ammonium chloride (2 ml) was added and the mixture was stirred for 10 minutes. The mixture was concentrated in vacuo and partitioned between water (50 ml) and DCM (50 ml). The aqueous layer was further extracted with DCM (2 × 50 ml) and the organic layers were combined and concentrated in vacuo. The crude product was purified by flash chromatography (0-100% EtOAc / heptane) to yield benzyl (3S, 4R) -3-{[(tert-butoxy) carbonyl] amino} -4-methoxypiperidine-1- The carboxylate EV-AT1700-001 [248 mg (52.9%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.29 minutes, M / z = 387 (M + 23).

  Tert-butyl N-[(3S, 4R) -4-methoxypiperidin-3-yl] carbamate EV-AT1701-001-step 7

  Stirring of benzyl (3S, 4R) -3-{[(tert-butoxy) carbonyl] amino} -4-methoxypiperidine-1-carboxylate (EV-AT1700-001, 235 mg, 0.64 mmol) / ethanol (10 ml) To the solution was added 10% Pd / C (34 mg, 0.03 mmol) under nitrogen. The reaction solution was placed under a hydrogen atmosphere and stirred at room temperature for 16 hours. The reaction mixture was filtered through Kieselguhr and the filtrate was washed with ethanol (20 ml). The filtrate was concentrated in vacuo to give tert-butyl N-[(3S, 4R) -4-methoxypiperidin-3-yl] carbamate EV-AT1701-001 [135 mg (68.0%)] as a colorless oil. Obtained. LCMS (Method D): retention time 0.79 min, M / z = 231- (M + 1).

  Tert-butyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate EV-AS-1581-001-Step 8

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-5-carboxylic acid (EV-AR3168) To a stirred solution of -002,150 mg, 0.43 mmol) / DMF (5 ml) was added HATU (197.6 mg, 0.52 mmol) followed by DIPEA (0.15 ml, 0.87 mmol). The mixture was stirred for 1 hour and then tert-butyl N-[(3S, 4R) -4-methoxypiperidin-3-yl] carbamate (EV-AT1701-001, 99.7 mg, 0.43 mmol) was added. . The reaction solution was stirred at room temperature for 16 hours. The solvent was removed in vacuo and the residue was partitioned between DCM (30 ml) and saturated aqueous sodium bicarbonate (30 ml). The aqueous layer was extracted with DCM (20 ml) and the organics were combined and washed with water (20 ml) and saturated aqueous sodium chloride solution (20 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give an orange oil which was purified by preparative HPLC (basic method) and tert-butyl N- [ (3S, 4R) -1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole -5-Carbonyl} -4-methoxypiperidin-3-yl] carbamate EV-AS1581-001 [205 mg (82.9%)] was obtained as a white crystalline solid. LCMS (Method A): retention time 3.36 minutes, M / z = 559 (M + 1).

  1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl}- 4-Methoxypiperidin-3-amine EV-AS1585-001- (EOAI3436357) I-62-Step 9

  tert-Butyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate (EV-AS1581-001, 30 mg, 0.05 mmol) / DCM (2 ml) in a stirred solution of trifluoroacetic acid (1 ml , 13.0 mmol). The mixture was stirred at room temperature for 3 hours. The solvent was removed under a nitrogen stream and the residue was lyophilized from acetonitrile: water (1: 1, 4 ml) to give 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2 , 3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-methoxypiperidin-3-aminetrifluoroacetic acid EV-AS1585-001, I- 62 [9.6 mg (96.3%)] was obtained as a white powder. LCMS (Method A): retention time 1.86 minutes, M / z = 459 (M + 1).

  tert-Butyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate EV-AS1581-002 and tert-butyl N-[(3R, 4S) -1- {2- [1- (cyclo Propylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate Chiral HPLC to obtain EV-AS1581-003—Step 10

  tert-Butyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate EV-AS1581-001 [90.4 mg] was dissolved in methanol and then purified by chiral HPLC (Method K). Tert-butyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate EV-AS1581-002 (absolute stereochemistry assigned arbitrarily) [36.2 mg] and tert-butyl N -[(3R, 4S) -1- {2- [ -(Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-methoxypiperidine-3- IL] carbamate EV-AS1581-003 (absolute stereochemistry assigned arbitrarily) [34.4 mg] was obtained.

EV-AS1581-002 chiral purity (UV, 254 nm): 100%, retention time: 7.58 minutes (Method L)
EV-AS1581-003 chiral purity (UV, 254 nm): 100%, retention time: 10.07 minutes (Method L)

  (3S, 4R) -1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole -5-carbonyl} -4-methoxypiperidin-3-amine EV-AS1592-001- (EOAI 3438020, absolute stereochemistry assigned arbitrarily) I-64-step 9

  Tert-butyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate (EV-AS1581-002, 36.2 mg, 0.07 mmol) was treated as in Step 9, Scheme 1. , (3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodi Azol-5-carbonyl} -4-methoxypiperidin-3-aminetrifluoroacetic acid EV-AS1592-001, I-64 [37.1 mg (64.6%)] was obtained as a white powder. LCMS (Method A): retention time 1.86 minutes, M / z = 459 (M + 1).

  (3R, 4S) -1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole -5-carbonyl} -4-methoxypiperidin-3-amine EV-AS1593-001- (EOAI 3437799, absolute stereochemistry arbitrarily assigned) I-65-Step 9

  Tert-butyl N-[(3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1 , 3-Benzodiazol-5-carbonyl} -4-methoxypiperidin-3-yl] carbamate (EV-AS1581-003, 34.4 mg, 0.06 mmol) was treated as in Step 9, Scheme 1. , (3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodi Azol-5-carbonyl} -4-methoxypiperidin-3-aminetrifluoroacetic acid EV-AS1593-001, I-65 [33.6 mg (94.3%)] was obtained as a white powder. LCMS (Method A): retention time 1.86 minutes, M / z = 459 (M + 1).

  5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridin-6-carbonyl] -2- [1- (cyclopropylmethyl) -5-phenyl-1H-pyrrol-2-yl ] -1-Methyl-1H-1,3-benzodiazole EV-AS5709-003 (EOAI3434977) I-116 and 5-[(3aS, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridine- 6-carbonyl] -2- [1- (cyclopropylmethyl) -5-phenyl-1H-pyrrol-2-yl] -1-methyl-1H-1,3-benzodiazole EV-AS5710-003 (EOAI 3434978) I-117 is described in Scheme 2 via the synthesis of methyl 5-phenyl-1H-pyrrole-2-carboxylate EV-AR0054-002 as described in Scheme 2.1. It was synthesized according to the method:

To a stirred solution of 5-phenyl-1H-pyrrole-2-carboxylic acid (CAS 6636-06-2,500 mg, 2.67 mmol) / toluene (10 ml) and methanol (3 ml) was added 2M (diazomethyl) (trimethyl) silane / hexane. (2 ml) was added and the mixture was stirred at room temperature for 30 minutes under nitrogen. To the reaction mixture was added acetic acid (1 ml) and the mixture was concentrated in vacuo to give methyl 5-phenyl-1H-pyrrole-2-carboxylate (EV-AR0054-002) [530 mg (99%)] as a pale yellow powder. Got as. LCMS (Method D): retention time 1.14 minutes, M / z = 202 (M + 1).

  3-[(2- {5-[(3R) -3-Aminopiperidin-1-carbonyl] -1-methyl-1H-1,3-benzodiazol-2-yl} -1H-pyrrolo [2,3 -B] pyridin-1-yl) methyl] -1-methylcyclobutan-1-ol EV-AU7275-001- (EOAI34555096) I-110 was converted to ethyl 1-[(3-hydroxy- 3-Methylcyclobutyl) methyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxylate Synthesized according to the method described in Scheme 1 via synthesis of EV-AU7265-001:

3- (Hydroxymethyl) -1-methylcyclobutan-1-ol EV-AU7264-001-Step 1
To a stirred solution of 3-hydroxy-3-methylcyclobutanecarboxylic acid (CAS16286-86-5, 950 mg, 7.30 mmol) / THF (30 ml) was added BH ₃ · Me ₂ S (4.96 ml, 9.93 mmol) − Add dropwise at 78 ° C. The reaction solution was warmed to room temperature and stirred for 16 hours. The reaction solution was quenched with anhydrous MeOH (20 ml). The resulting mixture was dried under reduced pressure to give 3- (hydroxymethyl) -1-methylcyclobutan-1-ol EV-AU7264-001 [200 mg (23.2%)] as a colorless oil.

  1-[(3-Hydroxy-3-methylcyclobutyl) methyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AU7265-001-Step 2

  Under a nitrogen atmosphere, a triphenylphosphine (1.39 g, 5.26 mmol) / THF (10 ml) solution was added to a stirred solution of DIAD (1.10 ml, 5.26 mmol) / dry THF (10 ml) at −20 ° C. added. The reaction mixture was stirred for 30 minutes and then a solution of ethyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate (CAS 221675-35-0,500 mg, 2.63 mmol) / THF (10 ml) was added. . Stirring was continued for an additional 30 minutes at -20 ° C. After this, a solution of 3- (hydroxymethyl) -1-methylcyclobutan-1-ol (EV-AU7264-001, [458 mg, 3.94 mmol]) / THF (5 ml) was added dropwise at −20 ° C. The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The mixture is concentrated in vacuo and the residue is purified by flash chromatography (5-80% EtOAc / heptane) to give ethyl 1-[(3-hydroxy-3-methylcyclobutyl) methyl] -1H-pyrrolo. [2,3-b] pyridine-2-carboxylate EV-AU7265-001 [200 mg (23.2%)] was obtained as a yellow oil. LCMS (Method D): retention time 1.10 minutes, M / z = 289 (M + 1).

N- (2- {5-[(3R) -3-aminopiperidin-1-carbonyl] -1-methyl-1H-1,3-benzodiazol-2-yl} -1- (cyclopropylmethyl)- 1H-pyrrolo [2,3-b] pyridin-4-yl) acetamide EV-AT0096-001- (EOAI3447170) I-69 was synthesized according to the method described in Scheme 2 tert-butyl N-[(3R) -1- {2- [4-Bromo-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole- 5-carbonyl} piperidin-3-yl] carbamate tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl)-] obtained according to scheme 2.3 starting from EV-AT0086-001 4-acetamido-1H-pi [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate Boc-deprotection of EV-AT0094-001 Obtained by:

Tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -4-[(diphenylmethylidene) amino] -1H-pyrrolo [2,3-b] pyridin-2-yl ] -1-Methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AT0090-001-Step 1

Tert-Butyl N-[(3R) -1- {2- [4-bromo-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-ene in toluene (4.0 ml) Yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate (EV-AT0086-001, 150 mg, 0.24 mmol), 1,1-diphenylmethanimine (53 .2 mg, 0.29 mmol), Pd ₂ dba ₃ (5.6 mg, 0.006 mmol), BINAP (11.4 mg, 0.018 mmol) and Cs ₂ CO ₃ (111.5 mg, 0.34 mmol) The mixture was stirred at 100 ° C. for 5 hours in a sealed tube. The reaction solution was cooled to room temperature and filtered through Kieselguhr washed with EtOAc. The filtrate is evaporated to dryness and the remaining residue is dissolved in DMSO and purified by preparative HPLC (basic method) to give tert-butyl N-[(3R) -1- {2- [1- (Cyclopropylmethyl) -4-[(diphenylmethylidene) amino] -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole -5-Carbonyl} piperidin-3-yl] carbamate EV-AT0090-001 [128 mg (73.2%)] was obtained as a yellow solid. LCMS (Method D): retention time 1.38 minutes, M / z = 708 (M + 1).

  Tert-butyl N-[(3R) -1- {2- [4-amino-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AT0092-001-Step 2

  Hydroxylamine hydrochloride (1: 1) (58.3 mg, 0.839 mmol) and NaOAc (90 mg, 1.09 mmol) were added to tert-butyl N-[(3R) -1- {2- [1- (cyclopropyl Methyl) -4-[(diphenylmethylidene) amino] -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidine To a suspension of -3-yl] carbamate (EV-AT0090-001, 120 mg, 0.168 mmol) / MeOH (8 ml). The resulting mixture was stirred at room temperature for 16 hours. The solvent is removed in vacuo and the remaining residue is dissolved in DMSO and purified by preparative HPLC (basic method) to give tert-butyl N-[(3R) -1- {2- [ 4-Amino-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3 -Il] carbamate EV-AT0092-001 [86 mg (94.3%)] was obtained as a white solid. LCMS (Method D): retention time 0.95 min, M / z = 544 (M + 1).

  Tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -4-acetamido-1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AT0094-001-Step 3

  tert-Butyl N-[(3R) -1- {2- [4-amino-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate (EV-AT0092-001,80 mg, 0.15 mmol) / DCM (4 ml) in acetic anhydride (18 mg, 0.18 mmol). ) And triethylamine (30 mg, 0.29 mmol) were added at room temperature and the reaction solution was stirred at room temperature for 16 hours and then at 50 ° C. for 24 hours. The solvent is removed in vacuo and the remaining residue is dissolved in DMSO and purified by preparative HPLC (basic method) to give tert-butyl N-[(3R) -1- {2- [ 1- (cyclopropylmethyl) -4-acetamido-1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidine-3 -Il] carbamate EV-AT0094-001 [82 mg (95.1%)] was obtained as a white foam. LCMS (Method D): retention time 1.09 minutes, M / z = 586 (M + 1).

  Tert-Butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- described in Scheme 2.4 (Hydroxymethyl) -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate Through the synthesis of EV-AV9641-002 according to the method described in Scheme 2 {5 -[(3R) -3-Aminopiperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H- 1,3-Benzodiazol-7-yl} methanol EV-AV 9647-001 (EOAI 3455579) I-128 was synthesized.

  Tert-Butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- described in Scheme 2.4 Through the synthesis of methanesulfonyl-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9652-001, (3R) -1- {2- [1 -(Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methanesulfonyl-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3 -Amine EV-AV9654-001 (EOAI3455786) I-131 was synthesized according to the method described in Scheme 2:

  Tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (hydroxymethyl) -1- Methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9641-002-step 1

  tert-Butyl N-[(3R) -1- {7-bromo-2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate (EV-AV9639-001, 50 mg, 0.08 mmol) / anhydrous THF (5.0 ml) at −78 ° C. N-Butyllithium (1.6M in hexane, 0.11 ml, 0.17 mmol) was added dropwise. The reaction solution was stirred at −78 ° C. for 10 min and anhydrous DMF (0.01 ml, 0.16 mmol) was added in one portion. The reaction mixture was stirred for 10 minutes at -78 ° C and allowed to warm to room temperature over 1 hour. The reaction mixture was then cooled to 0 ° C. and saturated aqueous ammonium chloride solution (1 ml) was added. The biphasic mixture was stirred for 30 minutes before the layers were separated. The aqueous phase was extracted again with EtOAc (2 × 3 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was dissolved in methanol (5 ml) at 0 ° C. and sodium borohydride (6 mg, 0.16 mmol) was added. The reaction mixture was stirred for 16 hours at room temperature, quenched by the addition of water (2 ml) and concentrated in vacuo. The residue was partitioned between EtOAc (5 ml) and water (2 ml), the aqueous layer was further extracted with EtOAc (2 × 2 ml) and the combined organic layers were dried over sodium sulfate and filtered. And concentrated in vacuo. The crude product was purified by preparative HPLC (basic method) and tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3- b] pyridin-2-yl] -7- (hydroxymethyl) -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9641-002 [20 mg (43 %)] Was obtained as an off-white powder. LCMS (Method D): retention time 1.10 minutes, M / z = 559 (M + 1).

  Tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methanesulfonyl-1-methyl- 1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9652-001-Step 2

  To the pressure tube, copper (II) triflate (45 mg, 0.12 mmol), sodium methanesulfonate (25 mg, 0.25 mmol) and N, N′-dimethylethane-1,2-diamine (0.03 ml, 0.26 mmol). ) And DMSO (2.0 ml) were added under a nitrogen atmosphere. The deep blue reaction was stirred at room temperature for 5 minutes to give tert-butyl N-[(3R) -1- {7-bromo-2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b ] Pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate (EV-AV9639-001, 75 mg, 0.12 mmol) was added. The vessel was sealed and heated at 120 ° C. for 2 hours. The cooled reaction mixture was diluted with water (15 ml) and extracted with EtOAc (2 × 10 ml). The organic layers were combined, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC (acidic method) and tert-butyl N-[(3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b ] Pyridin-2-yl] -7-methanesulfonyl-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9652-001 [20 mg (27%)] Was obtained as an off-white powder. LCMS (Method D): 1.18 min, M / z = 607 (M + 1).

2- {1-Ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6 -Carbonyl} -1H-1,3-benzodiazole EV-AR0076-002 (EOAI3428370) I-13

  Methyl 3-methoxy-4- (methylamino) -5-nitrobenzoate EV-AR0065-002-step 1

To a stirred solution of methyl 4-chloro-3-methoxy-5-nitrobenzoate (CAS 66033-09-8, 2.00 g, 8.14 mmol) / DMF (10 ml) was added K ₂ CO ₃ (99%, 1.37 g, 9.81 mmol) was added. To this solution was added methanamine hydrochloride (1: 1) (0.62 g, 9.18 mmol) and the mixture was stirred in a sealed tube at 80 ° C. for 16 hours under nitrogen. The crude product of the reaction was concentrated in vacuo and partitioned between DCM (100 ml) and water (10 ml). The organic layer was further washed with water (2 × 10 ml) and saturated aqueous sodium chloride solution (10 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give an orange powder that was purified by flash chromatography (15-40% EtOAc / heptane) to give methyl 3- Methoxy-4- (methylamino) -5-nitrobenzoate EV-AR0065-002 [1.49 g (76%)] was obtained as an orange powder. LCMS (Method D): retention time 1.13 minutes, M / z = 241- (M + 1).

  Methyl 3-amino-5-methoxy-4- (methylamino) benzoate EV-AR0068-002-step 2

  To a stirred solution of methyl 3-methoxy-4- (methylamino) -5-nitrobenzoate (EV-AR0065-002, 1.49 g, 6.20 mmol) / ethanol (100 ml) was added 10% Pd / C (0.18 g, 0.17 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction mixture was filtered through Kieselguhr and the filtrate was washed with methanol (150 ml). The filtrate was concentrated in vacuo to give methyl 3-amino-5-methoxy-4- (methylamino) benzoate EV-AR0068-002 [1.21 g (89%)] as a pale purple powder. LCMS (Method D): retention time 0.63 minutes, M / z = 211− (M + 1).

  Ethyl 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AQ1957-001-Step 3

  Sodium hydride (60%, 59 mg, 1.47 mmol) was added to ethyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate (CAS 221675-35-0, 200 mg, 1.05 mmol) / DMF (5 ml). To the stirred suspension of was added dropwise at room temperature. The mixture was stirred for 20 minutes and then ethyl iodide (197 mg, 1.26 mmol) was added. The reaction mixture was stirred for 20 hours. The mixture was partitioned between EtOAc (20 ml) and water (20 ml). The aqueous layer was further extracted with EtOAc (1 × 20 ml) and the combined organic layers were washed with water (20 ml) and evaporated to dryness. The crude product was purified by flash chromatography (0-50% EtOAc / heptane) to give ethyl 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AQ1957-001 [135 mg. (57.1%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.18 minutes, M / z = 219 (M + 1).

  1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AQ1960-001-step 4

  To a stirred solution of ethyl 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AQ1957-001, 135 mg, 0.62 mmol) / THF (2 ml) was added lithium hydroxide (74 mg, 3.09 mmol) / water (2 ml) was added. The mixture was stirred at 50 ° C. for 2.5 hours, acidified with 1M HCl (3 ml) and extracted with DCM (2 × 5 ml). The combined organics were washed with water and evaporated to dryness to give 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AQ1960-001 [120 mg (99%)] as a white solid. Obtained. LCMS (Method D): retention time 0.92 min, M / z = 191- (M + 1).

  Methyl 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AR0070 -003-Step 5

  To a stirred solution of 1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AQ1960-001, 120 mg, 0.63 mmol) / DMF (2 ml) was added DIPEA (116 μl, 0.70 mmol). ), Then HATU (236 mg, 0.62 mmol) was added and the resulting mixture was stirred at room temperature for 15 minutes. Methyl 3-amino-5-methoxy-4- (methylamino) benzoate (EV-AR0068-002, 148 mg, 0.70 mmol) was obtained and the resulting mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated in vacuo, dissolved in acetic acid (3 ml) and stirred at 70 ° C. for 16 hours. The solvent was removed in vacuo and the residual material was purified by flash chromatography (25-40% EtOAc / heptane) to give methyl 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridine-2. -Il} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AR0070-003 [150 mg (63%)] was obtained as a white powder. LCMS (Method D): retention time 1.18 minutes, M / z = 365 (M + 1).

  2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid EV-AR0072- 002-Step 6

  Methyl 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate (EV- To a stirred solution of AR0070-003, 150 mg, 0.41 mmol) / THF (3 ml) was added a solution of lithium hydroxide (30 mg, 1.25 mmol) / water (3 ml) and the mixture was stirred at room temperature for 16 hours. . The reaction mixture was concentrated in vacuo, taken up in water (5 ml) and acidified with 5N HCl (0.5 ml) with stirring. The resulting suspension was stirred for 10 minutes and then the precipitate was collected by vacuum filtration to give 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7- Methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid EV-AR0072-002 [130 mg (89%)] was obtained as a white powder. LCMS (Method D): retention time 1.03 min, M / z = 351- (M + 1).

  Tert-butyl 6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 Carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0074-002-Step 7

  2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid (EV-AR0072 To a stirred solution of −002,130 mg, 0.37 mmol) / 2: 1 DMSO / MeCN (4.5 ml) was added DIPEA (65 μl, 0.39 mmol) and HATU (148 mg, 0.39 mmol). The resulting mixture was stirred at room temperature for 15 minutes and then tert-butyl octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (CAS 949559-11-9,88 mg, 0.39 mmol) was added. The mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with 3: 2 MeCN / water (1.5 ml) and purified by preparative HPLC (basic method) to give tert-butyl 6- (2- {1-ethyl-1H- Pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] Pyridine-1-carboxylate EV-AR0074-002 [142 mg (81%)] was obtained as a white powder. LCMS (Method A): retention time 3.41 min, M / z = 559 (M + 1).

  2- {1-Ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6 -Carbonyl} -1H-1,3-benzodiazole EV-AR0076-002 (EOAI3428370) I-13-step 8

  tert-butyl 6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 To a stirred solution of carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AR0074-002, 20 mg, 0.04 mmol) / methanol (1 ml) 4M HCl / 1,4 -Dioxane (0.5 ml) was added and the resulting solution was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuo and the residue was lyophilized from water (4 ml) to give 2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy- 1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl} -1H-1,3-benzodiazole hydrochloride EV-AR0076-002, I-13 [13.4 mg (75%)] was obtained as a yellow solid. LCMS (Method A): retention time 1.80 minutes, M / z = 459 (M + 1).

  tert-Butyl (3aR, 7aS) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0090-001 and tert-butyl (3aS, 7aR) -6- (2- {1- Ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2, 3-c] Pyridine-1-carboxylate Chiral HPLC to obtain EV-AR0090-002—Step 9

  tert-butyl 6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 Carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0074-002 [107 mg] was dissolved in ethanol and then purified by chiral HPLC (Method E) to give tert -Butyl (3aR, 7aS) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] -pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0090-001- (absolute stereochemistry assigned arbitrarily) [46.6 mg (43 .6%)] and tert Butyl (3aS, 7aR) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] -pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzo Diazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AR0090-002 (absolute stereochemistry assigned arbitrarily) [36.8 mg (33.4 %)].

EV-AR0090-001 chiral purity (UV, 254 nm): 100%, retention time: 6.30 minutes (Method F)
EV-AR0090-002 chiral purity (UV, 254 nm): 97%, retention time: 9.96 minutes (Method F)

  5-[(3aS, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridin-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl } -7-Methoxy-1-methyl-1H-1,3-benzodiazole EV-AR0091-002 (EOAI 3342499, absolute stereochemistry arbitrarily assigned) I-3-Step 8

  Tert-butyl (3aR, 7aS) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AR0090-001, 46 mg, 0.08 mmol) as in step 8, scheme 2. 5-[(3aS, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b] pyridine -2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole hydrochloride EV-AR0091-002, I-3 [40 mg (97%)] was obtained as an orange powder. LCMS (Method A): retention time 1.83 minutes, M / z = 459 (M + 1).

  5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridin-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl } -7-Methoxy-1-methyl-1H-1,3-benzodiazole EV-AR0092-002 (EOAI 3432500, absolute stereochemistry assigned arbitrarily) I-14-Step 8

  Tert-butyl (3aS, 7aR) -6- (2- {1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl) -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AR0090-002, 36.8 mg, 0.07 mmol) was prepared according to step 8, scheme 2. 5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- {1-ethyl-1H-pyrrolo [2,3-b ] Pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole EV-AR0092-002, I-14 [31 mg (92.7%)] was obtained as an orange powder. . LCMS (Method A): retention time 1.82 minutes, M / z = 459 (M + 1).

Special Case for Scheme 3 Ethyl 1-[(1-methylcyclopropyl) methyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxylate described in Scheme 3.1 EV-AU3682-002 Through synthesis, (3S, 5S) -5-fluoro-1- (7-methoxy-1-methyl-2- {1-[(1-methylcyclopropyl) methyl] -1H-pyrrolo [2,3- b] Pyridin-2-yl} -1H-1,3-benzodiazole-5-carbonyl) piperidin-3-amine EV-AV3056-001- (EOAI34554078) I-93 was synthesized according to the method described in Scheme 2 did:

  To a solution of DIAD (0.44 ml, 2.10 mmol) / dry THF (5 ml) at −20 ° C. under nitrogen was added a solution of triphenylphosphine (557 mg, 2.10 mmol) / THF (5 ml). The reaction mixture was stirred for 30 minutes. To the reaction mixture was added a solution of ethyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate (CAS 394223-02-0, 200 mg, 1.05 mmol) / THF (7.5 ml) at −20 ° C. The reaction mixture was stirred at −20 ° C. for a further 30 minutes. (1-Methylcyclopropyl) methanol (CAS 2746-14-7, 0.15 ml, 1.58 mmol) was added dropwise at −20 ° C. and the reaction mixture was allowed to warm to room temperature and stirred for 1 hour 15 minutes. did. The reaction mixture was concentrated in vacuo and purified by flash column chromatography (0-100% EtOAc) to give ethyl 1-[(1-methylcyclopropyl) methyl] -1H-pyrrolo [2,3-b] pyridine. 2-Carboxylate EV-AU3682-002 [240 mg (87%)] was obtained as a yellow gum. LCMS (Method D): retention time 1.35 minutes, M / z = 259 (M + 1).

  (3R) -1- (2- {1-[(1R) -1-cyclopropylethyl] -1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H -1,3-benzodiazol-5-carbonyl) piperidin-3-amine EV-AV3097-001- (EOAI34554812) I-101 and (3R) -1- (2- {1-[(1S) -1- Cyclopropylethyl] -1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl) piperidin-3-amine EV -AV3098-001- (EOAI34554813) I-102 was converted to ethyl 1- (1-cyclopropylethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AU3682 as described in Scheme 3.2. -001 synthesis To, was synthesized according to the method described in Scheme 2:

  A solution of triphenylphosphine (557.18 mg, 2.1 mmol) / THF (5 ml) was added to a solution of DIAD (0.44 ml, 2.1 mmol) / dry THF (5 ml) at −20 ° C. under nitrogen. In addition, the reaction mixture was stirred for 30 minutes. To the reaction mixture was added a solution of ethyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate (CAS 394223-02-0, 200 mg, 1.05 mmol) / THF (7.5 ml) at −20 ° C. The reaction mixture was stirred at −20 ° C. for a further 30 minutes. 1-Cyclopropylethane-1-ol (CAS765-42-4, 0.15 mL, 1.58 mmol) was added dropwise at −20 ° C., the reaction mixture was allowed to warm to room temperature and stirred for 1 hour 15 minutes. . The reaction mixture was concentrated in vacuo and purified using flash column chromatography (0-100% EtOAc) to yield ethyl 1- (1-cyclopropylethyl) -1H-pyrrolo [2,3-b] pyridine- 2-carboxylate EV-AU3682-001 [205 mg (75%)] was obtained as a yellow gum. LCMS (Method D): retention time 1.35 minutes, M / z = 259 (M + 1).

  (3R) -1- {2- [1- (Cyclopropylmethyl) -6-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1 , 3-Benzodiazole-5-carbonyl} piperidin-3-amine EV-AV 4845-001- (EOAI 3454972) I-105 is converted to methyl 2- [1- (cyclopropylmethyl) -6 described in Scheme 3.3. Synthesis of -ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxylate EV-AV4834-002 Via the method described in Scheme 2:

Methyl 2- [6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazole -5-Carboxylate EV-AV4831-001 (synthesized according to Scheme 2, 150 mg, 0.31 mmol) / anhydrous THF (5 ml) in a solution of Fe (acac) ₃ (6 mg, 0.02 mmol) and NMP (150 [mu] l) Was added. A solution of ethylmagnesium bromide (0.9M in THF, 411 μl, 0.37 mmol) was added dropwise over 1 minute and the reaction mixture was stirred at room temperature for 3 hours. Additional Fe (acac) ₃ (6 mg, 0.02 mmol) and ethylmagnesium bromide (0.9 M in THF, 411 μl, 0.37 mmol) were added and the reaction mixture was stirred at room temperature for 16 hours. The reaction was quenched by the addition of 1M HCl (˜1 ml) and extracted with DCM (3 × 15 ml). The organic layers were combined, washed with brine (20 ml), dried over magnesium sulfate and concentrated in vacuo. The crude residue was purified by preparative HPLC (acidic method) to give methyl 2- [1- (cyclopropylmethyl) -6-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -1-Ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxylate EV-AV4834-002 [81 mg (55%)] was obtained as a colorless glass. LCMS (Method A): retention time 4.59 minutes, M / z = 433 (M + 1).

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 -Carbonyl} -5- (methylamino) piperidin-3-ol EV-AV4609-001- (EOAI3451154) I-77 was synthesized according to Scheme 3.4:

Scheme 3.4

  Rel-tert-butyl (3R, 5R) -3-hydroxy-5- (methylamino) piperidine-1-carboxylate EV-AV4600-001-Step 1. Note: All starting materials and products are trans-racemic.

  sodium hydride in a solution of rel-tert-butyl (3R, 5R) -3-amino-5-hydroxypiperidine-1-carboxylate hydrochloride (100 mg, 0.40 mmol) / anhydrous THF (5 ml) at 0 ° C. Was added (60%, 35 mg, 0.87 mmol) and the mixture was stirred at 0 ° C. for 30 min. To the reaction mixture was added methyl iodide (26 μl, 0.41 mmol) and the mixture was allowed to warm to room temperature and continued to stir for 16 hours. The reaction mixture was partitioned between ethyl acetate (20 ml) and water (20 ml). The aqueous extract is further washed with ethyl acetate (2 × 15 ml) and the organic extracts are combined, dried over sodium sulfate, filtered, concentrated in vacuo and rel-tert-butyl (3R, 5R) -3- Hydroxy-5- (methylamino) piperidine-1-carboxylate EV-AV4600-001 [91 mg (quantitative)] was obtained as a yellow oil. LCMS (Method D): retention time 0.23 min, M / z = 231 (M + 1).

  Rel-tert-butyl (3R, 5R) -3-{[(benzyloxy) carbonyl] (methyl) amino} -5-hydroxypiperidine-1-carboxylate EV-AV4602-001-step 2. Note: Starting material and product are trans-racemic.

    To a stirred solution of rel-tert-butyl (3R, 5R) -3-hydroxy-5- (methylamino) piperidine-1-carboxylate EV-AV4600-001- (91 mg, 0.40 mmol) / DCM (2 ml), DIPEA (103 μl, 0.59 mmol) was added followed by benzyl chloroformate (56 μl, 0.40 mmol). The reaction mixture was stirred for 1.5 hours, diluted with DCM (20 ml) and washed with water (15 ml). The organic extract was dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (0-100% EtOAc / heptane) to give rel-tert-butyl (3R, 5R) -3-{[(benzyloxy) carbonyl] (methyl) amino} -5. -Hydroxypiperidine-1-carboxylate EV-AV4602-001 [58 mg (18%)] was obtained as a clear oil. LCMS (Method D): retention time 1.12 minutes, M / z = 387 (M + Na).

Rel-benzyl N-[(3R, 5R) -5-hydroxypiperidin-3-yl] -N-methylcarbamate hydrochloride EV-AV4603-001-step 3
Note: Starting material and product are trans-racemic.

  rel-tert-butyl (3R, 5R) -3-{[(benzyloxy) carbonyl] (methyl) amino} -5-hydroxypiperidine-1-carboxylate EV-AV4602-001- (58 mg, 0.16 mmol) / To a solution of dioxane (1 ml) was added 4M HCl / dioxane (0.16 ml, 0.64 mmol) under nitrogen. The mixture was left at room temperature for 16 hours. To the reaction mixture was added methanol (0.5 ml) and 4M HCl / dioxane (0.16 ml, 0.64 mmol) and the mixture was allowed to stand at room temperature for 4 hours. The reaction mixture was then concentrated in vacuo to give rel-benzyl N-[(3R, 5R) -5-hydroxypiperidin-3-yl] -N-methylcarbamate hydrochloride EV-AV4603-001 [59 mg (43%)] Was obtained as an off-white solid. LCMS (Method D): retention time 0.76 min, M / z = 265 (M + 1).

  Benzyl N-[(3R, 5R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-benzodiazol-5-carbonyl} -5-hydroxypiperidin-3-yl] -N-methylcarbamate EV-AV 4604-001-Step 4

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid A solution of EV-AU3032-001 (synthesized according to Scheme 2, 60 mg, 0.16 mmol) / DMF (3 ml) was added to DIPEA (31 μl, 0.18 mmol), HATU (67 mg, 0.18 mmol) and rel-benzyl N- [(3R, 5R) -5-Hydroxypiperidin-3-yl] -N-methylcarbamate hydrochloride EV-AV4603-001- (48 mg, 0.16 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour. . The mixture was then partitioned between EtOAc (40 ml) and water (40 ml) and the organic extract was dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by preparative HPLC (basic method) to give benzyl N-[(3R, 5R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3 -B] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -5-hydroxypiperidin-3-yl] -N-methylcarbamate EV-AV4604 -001 [22 mg (22%)] was obtained as an off-white solid. LCMS (Method D): retention time 1.17 minutes, M / z = 623 (M + 1).

  Rel- (3R, 5R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -5- (methylamino) piperidin-3-ol hydrochloride EV-AV4609-001, I-77-Step 5. Note: Starting material and product are trans racemic.

  rel-benzyl N-[(3R, 5R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazol-5-carbonyl} -5-hydroxypiperidin-3-yl] -N-methylcarbamate EV-AV 4604-001 (22 mg, 0.04 mmol) / stirring solution of ethanol (1 ml) To the solution was added 10% Pd / C (2.3 mg, 0.001 mmol) under nitrogen and the resulting mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction mixture was filtered through a glass fiber sinter and then the filtrate was treated with 1.25 M HCl / ethanol (0.2 ml). The mixture was left at room temperature for 30 minutes, concentrated in vacuo, lyophilized and rel- (3R, 5R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2, 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -5- (methylamino) piperidin-3-ol hydrochloride EV-AV4609 -001, I-77 [10.2 mg (52%)] was obtained as an off-white powder. LCMS (Method A): retention time 1.93 minutes, M / z = 489 (M + 1).

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 -Carbonyl} -4- (difluoromethoxy) piperidin-3-amine EV-AV4627-001 (EOAI34552884) I-86 was synthesized according to Scheme 3.5:

  Rac-tert-butyl (3R, 4R) -3-{[(benzyloxy) carbonyl] amino} -4-hydroxypiperidine-1-carboxylate EV-AV3800-001-step 1- (trans racemate)

  To a solution of rac-tert-butyl (3R, 4R) -3-amino-4-hydroxypiperidine-1-carboxylate (CAS 1523530-23-5, 750 mg, 3.47 mmol) / DCM (10 ml) at 0 ° C., Triethylamine (1.45 ml, 10.4 mmol) and benzyl chloroformate (0.59 ml, 4.16 mmol) were added. The reaction mixture was stirred at 0 ° C. for 15 minutes, then allowed to warm to room temperature and stirred for an additional 4 hours. The reaction mixture was quenched with water (10 ml) and extracted with DCM (3 × 10 ml). The combined organic fractions were dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (20-80% EtOAc / heptane) to give rac-tert-butyl (3R, 4R) -3-{[(benzyloxy) carbonyl] amino} -4-hydroxypiperidine. -1-Carboxylate EV-AV3800-001 [615 mg (52%)] was obtained as a white solid (trans-racemate). LCMS (Method D): retention time 1.08 min, M / z = 373 (M + Na).

  rac-benzyl N-[(3R, 4R) -4-hydroxypiperidin-3-yl] carbamate EV-AV3801-001-step 2 (trans racemate)

  Rac-tert-butyl (3R, 4R) -3-{[(benzyloxy) carbonyl] amino} -4-hydroxypiperidine-1-carboxylate EV-AV3800-001 (610 mg, 1.74 mmol) was added to dioxane (dioxane). In 4M, 8.7 ml) and stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo to give rac-benzyl N-[(3R, 4R) -4-hydroxypiperidin-3-yl] carbamate EV-AV3801-001 [408 mg (82%)] as a white solid. LCMS (Method D): retention time 0.48 min, M / z = 251- (M + 1).

  rac-benzyl N-[(3R, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazol-5-carbonyl} -4-hydroxypiperidin-3-yl] carbamate EV-AV3805-001-step 3 (trans racemate)

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5 -To a solution of carboxylic acid EV-AV3032-001 (535 mg, 1.42 mmol) / DMF (5 ml) was added HATU (595 mg, 1.57 mmol) and DIPEA (0.59 ml, 3.56 mmol) and the reaction mixture was And stirred at room temperature for 30 minutes. rac-Benzyl N-[(3R, 4R) -4-hydroxypiperidin-3-yl] carbamate EV-AV3801-001 (synthesized according to Scheme 2, 408 mg, 1.42 mmol) was then added and the reaction mixture was The mixture was further stirred at room temperature for 2 hours. The reaction mixture was then diluted with EtOAc (10 ml) and washed with water (3 × 10 ml) and saturated aqueous sodium chloride solution (10 ml). The organic fraction was then dried (sodium sulfate), filtered and concentrated in vacuo to give rac-benzyl N-[(3R, 4R) -1- {2- [1- (cyclopropylmethyl) -1H. -Pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-hydroxypiperidin-3-yl] carbamate EV -AV3805-001 [775 mg (79%)] was obtained as an orange solid. LCMS (Method D): retention time 1.12 minutes, M / z = 609 (M + 1).

  rac- (3R, 4S) -3-{[(Benzyloxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl ] -7-Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-4-yl benzoate EV-AV3814-001-Step 4 (cis racemate)

  To a solution of triphenylphosphine (411 mg, 1.55 mmol) / THF (1 mol) at 0 ° C. was added DIAD (325 μl, 1.55 mmol). The reaction mixture was stirred for 5 minutes and rac-benzyl N-[(3R, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2- Yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-hydroxypiperidin-3-yl] carbamate EV-AV3805-001 (675 mg, 1.11 mmol) and Benzoic acid (190 mg, 1.55 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours and then diluted with EtOAc (15 ml). The mixture was washed with water (10 ml) and saturated aqueous sodium chloride solution (10 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (50-100% EtOAc / heptane) to give rac- (3R, 4S) -3-{[(benzyloxy) carbonyl] amino} -1- {2- [1 -(Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-4- IL-benzoate EV-AV3814-001 [743 mg (49%)] was obtained. LCMS (Method D): retention time 1.36 minutes, M / z = 713 (M + 1).

  rac-benzyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazole-5-carbonyl} -4-hydroxypiperidin-3-yl] carbamate EV-AV3819-001-Step 5 (cis racemate)

rac- (3R, 4S) -3-{[(Benzyloxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl ] -7-Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-4-yl benzoate EV-AV3814-001- (0.74 g, 0.54 mmol) / ethanol (10 ml) To a solution of and water (5 ml) was added K ₂ CO ₃ (0.11 g, 0.81 mmol) and the solution was stirred at room temperature for 3 hours. The reaction mixture was then concentrated in vacuo and the crude residue was dissolved in DCM (10 ml) and washed with water (10 ml). The organic layer was then washed with saturated aqueous sodium chloride solution (10 ml), dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography (0-20% methanol / EtOAc) and rac-benzyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H- Pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-hydroxypiperidin-3-yl] carbamate EV- AV3819-001 [214 mg (64%)] was obtained as a white solid. LCMS (Method D): retention time 1.14 minutes, M / z = 609 (M + 1).

  rac-benzyl N-[(3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazol-5-carbonyl} -4- (difluoromethoxy) piperidin-3-yl] carbamate EV-AV4626-001-step 6 (cis racemate)

  Rac-Benzyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] in acetonitrile (2 ml) in a pressure tube ] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-hydroxypiperidin-3-yl] carbamate (EV-AV3819-001, 214 mg, 0.35 mmol) and iodine To a stirred suspension of copper (I) chloride (13.4 mg, 0.070 mmol) was added 2,2-difluoro-2- (fluorosulfonyl) acetic acid (125 mg, 0.70 mmol). The vessel was sealed and the reaction mixture was stirred at 80 ° C. for 2 hours. To the cooled reaction mixture was added 2,2-difluoro-2- (fluorosulfonyl) acetic acid (125 mg, 0.70 mmol), the vessel was sealed and the reaction mixture was stirred at 80 ° C. for an additional 2 hours. The reaction mixture is cooled again, 2,2-difluoro-2- (fluorosulfonyl) acetic acid (125 mg, 0.70 mmol) is added, the vessel is sealed and the reaction mixture is stirred at 80 ° C. for a further 2 hours. did. The reaction mixture is then concentrated in vacuo and the residue is purified by flash chromatography (20-100% EtOAc / heptane) followed by preparative HPLC (basic method) to give rac-benzyl N-[( 3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl} -4- (difluoromethoxy) piperidin-3-yl] carbamate EV-AV4626-001 [33 mg (14%)] was obtained as a white solid. LCMS (Method D): retention time 1.27 minutes, M / z = 659 (M + 1).

  rac- (3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -4- (difluoromethoxy) piperidin-3-amine EV-AV4627-001- (EOAI34552884) I-86-Step 7 (cis racemic)

  rac-benzyl N-[(3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazol-5-carbonyl} -4- (difluoromethoxy) piperidin-3-yl] carbamate (EV-AV4626-001, 33 mg, 0.05 mmol) / stirring solution of ethanol (2 ml) To the mixture was added 10% Pd / C (3.2 mg, 0.002 mmol) under nitrogen and the resulting mixture was stirred at room temperature for 5 hours under hydrogen atmosphere. The reaction mixture was filtered through a glass fiber sinter and then the filtrate was diluted with 1.25 M HCl / ethanol (0.3 ml). The mixture was left at room temperature for 30 minutes, concentrated in vacuo, lyophilized and rac- (3R, 4S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2, 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -4- (difluoromethoxy) piperidin-3-amine EV-AV4627-001 -(I-86) [28.2 mg (98%)] was obtained as an off-white powder. LCMS (Method A): retention time 2.20 minutes, M / z = 524 (M + 1).

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 -Carbonyl} -4-fluoropiperidin-3-amine EV-AV3807-001- (EOAI3451007) I-75 was synthesized according to the method described in Scheme 3.6:

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 -Carbonyl} -4-fluoropiperidin-3-yl methanesulfonate EV-AV 3803-001-step 1

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 -Carbonyl} -4-fluoropiperidin-3-ol (synthesized according to EV-AU3293-001, scheme 2, 570 mg, 1.19 mmol) / DCM (10 ml) at 0 ° C. with triethylamine (0.25 ml). , 1.79 mmol) and mesyl chloride (0.11 ml, 1.43 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. Water (5 ml) is added to the mixture and the organic layer is collected, dried over sodium sulfate, concentrated in vacuo, and 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2, 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -4-fluoropiperidin-3-yl methanesulfonate EV-AV-3803 001 [663 mg (quantitative)] was obtained. LCMS (Method D): retention time 1.14 minutes, M / z = 556 (M + 1).

  5- (3-Azido-4-fluoropiperidine-1-carbonyl) -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazole EV-AV 3804-001-step 2

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 To a stirred solution of -carbonyl} -4-fluoropiperidin-3-yl methanesulfonate (EV-AV-3803-001, 610 mg, 1.10 mmol) / DMSO (10 ml) was added sodium azide (285 mg, 4.39 mmol). added. The reaction mixture was stirred at 120 ° C. for 14 hours. The reaction mixture was cooled to room temperature, then diluted with EtOAc (20 ml), washed with water (3 × 20 ml) and then with saturated aqueous sodium chloride solution (20 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (50-100% EtOAc / heptane) to give 5- (3-azido-4-fluoropiperidine-1-carbonyl) -2- [1- (cyclopropylmethyl)- 1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole EV-AU3804-001 [191 mg (34.6%)] in orange Obtained as a colored solid. LCMS (Method D): retention time 1.19 minutes, M / z = 503 (M + 1).

  1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5 -Carbonyl} -4-fluoropiperidin-3-amine EV-AV3807-001 (EOAI34510007) I-75-step 3

  5- (3-Azido-4-fluoropiperidine-1-carbonyl) -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -To a stirred solution of methyl-1H-1,3-benzodiazole (EV-AU 3804-001, 191 mg, 0.38 mmol) / EtOAc (5 ml) under nitrogen, 10% Pd / C (81 mg, 0.8 04 mmol) was added. The mixture was left under a hydrogen atmosphere and stirred at room temperature for 12 hours. The mixture was filtered through a glass fiber filter and the filtrate was washed with methanol. The filtrate is concentrated in vacuo and the residue is purified by preparative HPLC (basic method) to give 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b]. Pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -4-fluoropiperidin-3-amine EV-AV3807-001- (I-75) [ 80 mg (44.2%)] was obtained as a white solid. LCMS (Method A): retention time 2.01 min, M / z = 477 (M + 1).

  5-Amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzo Diazole-5-carbonyl} piperidine-3-carbonitrile EV-AV4621-001 (EOAI3452077) I-82 was converted to methyl 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b ] Pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -5-hydroxypiperidine-3-carboxylate EV-AT8698-001 via scheme Synthesized according to the method described in 3.7. This was prepared according to the method described in Scheme 3, Step 7 using methyl 5-hydroxypiperidine-3-carboxylate. Methyl 5-hydroxypiperidine-3-carboxylate is prepared from 1-benzyl 3-methyl 5-hydroxypiperidine-1,3-dicarboxylate (CAS 1095010-45-9) according to the method described in Scheme 3, Step 7. did.

  Methyl 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- 5-carbonyl} -5- (methanesulfonyloxy) piperidine-3-carboxylate EV-AT8699-001-step 1

  Methyl 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- To a stirred solution of 5-carbonyl} -5-hydroxypiperidine-3-carboxylate (EV-AT8698-001, 275 mg, 0.53 mmol) / DCM (8 ml) at 0 ° C., triethylamine (0.11 ml, 0.80 mmol). ) And mesyl chloride (49 μl, 0.64 mmol) were added. The mixture was stirred for 2 hours with ice cooling. Further mesyl chloride (25 μl, 0.32 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours. Water (5 ml) was added to the mixture and the organic layer was collected, dried over sodium sulfate, concentrated in vacuo, and methyl 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [ 2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -5- (methanesulfonyloxy) piperidine-3-carboxylate EV -AT8699-001 [316 mg (quantitative)] was obtained as a yellow oil. LCMS (Method D): retention time 1.12 minutes, M / z = 596 (M + 1).

  Methyl 5-azido-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl} piperidine-3-carboxylate EV-AV4601-001-Step 2

  Methyl 1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- To a stirred solution of 5-carbonyl} -5- (methanesulfonyloxy) piperidine-3-carboxylate (EV-AT8699-001, 316 mg, 0.53 mmol) / DMSO (2 ml) was added sodium azide (86 mg, 1.33 mmol). ) Was added. The resulting mixture was stirred at 90 ° C. for 16 hours. The reaction mixture was cooled to room temperature and partitioned between EtOAc (40 ml) and water (30 ml). The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude residue was purified by flash chromatography (50-100% EtOAc / heptane) to give methyl 5-azido-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b. ] Pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidine-3-carboxylate EV-AV4601-001 [141 mg (43.1%)] Was obtained as an off-white solid. LCMS (Method D): retention time 1.19 minutes, M / z = 543 (M + 1).

  Methyl 5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3- Benzodiazole-5-carbonyl} piperidine-3-carboxylate EV-AV4605-001-Step 3

  Methyl 5-azido-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3- To a stirred solution of benzodiazole-5-carbonyl} piperidine-3-carboxylate (EV-AV4601-001, 131 mg, 0.24 mmol) / ethanol under nitrogen, 10% Pd / C (0.15 g, 0 .01 mmol) was added. The reaction mixture was placed under a hydrogen atmosphere and stirred at room temperature for 16 hours. The reaction mixture is filtered through a glass fiber filter and the filtrate is concentrated in vacuo to give methyl 5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine- 2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylate EV-AV 4605-001 [98 mg (46.0%)] as a yellow oil Obtained as a thing. LCMS (Method D): retention time 0.92 min, M / z = 517 (M + 1).

  Methyl 5-{[(tert-butoxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy- 1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylate EV-AV4610-001-step 4

  Methyl 5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3- To a stirred solution of benzodiazole-5-carbonyl} piperidine-3-carboxylate (EV-AV 4605-001, 77 mg, 0.15 mmol) / dioxane (1 ml) was added saturated sodium bicarbonate solution (1 ml) and anhydrous Boc (39 mg , 0.18 mmol) was added. The resulting mixture was stirred at room temperature for 1 hour. The mixture was partitioned between EtOAc (20 ml) and water (20 ml). The aqueous layer was extracted with EtOAc (15 ml) and the combined organic layers were dried over sodium sulfate and concentrated in vacuo. The crude residue was purified by flash chromatography (0-100% EtOAc / heptane) to give methyl 5-{[(tert-butoxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylate EV-AV4610- 001 [62 mg (63.2%)] was obtained as an off-white powder. LCMS (Method D): retention time 1.20 minutes, M / z = 617 (M + 1).

  5-{[(tert-butoxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylic acid EV-AV4611-001-step 5

  Methyl 5-{[(tert-butoxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy- To a stirred solution of 1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylate (EV-AV4610-001, 62 mg, 0.10 mmol) / methanol (1 ml) was added 1M hydroxylated. Aqueous sodium solution (0.5 ml) was added. The resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo to remove methanol. The resulting aqueous solution was acidified to pH˜4-5 with 1M aqueous hydrochloric acid until a precipitate formed. The solid was filtered off and dried under vacuum to give 5-{[(tert-butoxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b]. Pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylic acid EV-AV4611-001 [58 mg (89.4%)]. Obtained as an off-white powder. LCMS (Method D): retention time 1.13 minutes, M / z = 603 (M + 1).

  Tert-butyl N- (5-carbamoyl-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl) carbamate EV-AV4612-001-step 6

  5-{[(tert-butoxy) carbonyl] amino} -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carboxylic acid (EV-AV4611-001, 58 mg, 0.96 mmol) / DMF (1 ml) in DIPEA (34 μl, 0.19 mmol), HATU (46 mg, 0.12 mmol) and ammonium chloride (10 mg, 0.19 mmol) were added. The resulting mixture was stirred at room temperature for 1 hour. The mixture was partitioned between EtOAc (40 ml) and water (40 ml). The aqueous layer was further extracted with EtOAc (40 ml) and the combined organic layers were dried over sodium sulfate, concentrated in vacuo and tert-butyl N- (5-carbamoyl-1- {2- [1- ( Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl) Carbamate EV-AV4612-001 [65 mg (97.7%)] was obtained as a yellow oil. LCMS (Method D): retention time 1.09 minutes, M / z = 602 (M + 1).

  Tert-butyl N- (5-cyano-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl) carbamate EV-AV4615-004-step 7

  tert-Butyl N- (5-carbamoyl-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H 1,3-benzodiazol-5-carbonyl} piperidin-3-yl) carbamate (EV-AV4612-001, 65 mg, 0.10 mmol) / anhydrous dioxane (1 ml) in a stirred solution of triethylamine (44.3 μl, 0.32 mmol) and trifluoroacetic anhydride (30 μl, 0.22 mmol) were added. The reaction mixture was stirred at room temperature for 5 hours. The mixture was treated again with triethylamine (44.3 μl, 0.32 mmol) and trifluoroacetic anhydride (30 μl, 0.22 mmol) and stirred at room temperature for 16 hours. The mixture was treated again with triethylamine (44.3 μl, 0.32 mmol) and trifluoroacetic anhydride (30 μl, 0.22 mmol) and stirred at room temperature for 2 hours. The mixture was treated again with (44.3 μl, 0.32 mmol) and trifluoroacetic anhydride (30 μl, 0.22 mmol) and stirred for 2 hours. The mixture was treated again with triethylamine (44.3 μl, 0.32 mmol) and trifluoroacetic anhydride (30 μl, 0.22 mmol) and stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc (20 ml) and saturated aqueous ammonium chloride (20 ml). The aqueous layer was extracted with EtOAc (20 ml) and the combined organic layers were dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash chromatography (0-100% EtOAc / heptane) and purified separately by preparative HPLC (basic method) to give two batches of product, tert-butyl N -[(3R, 5S) -5-cyano-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV4615-003 [9 mg (14.3%)] was obtained as an off-white powder. It was arbitrarily partitioned as the racemic cis-diastereomer. LCMS (Method D): retention time 1.21 min, M / z = 584 (M + 1). tert-Butyl N-[(3R, 5S) -5-cyano-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-Methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV4615-004 [11 mg (17.5%)] was obtained as an off-white powder. Arbitrarily distributed as racemic trans-diastereomers. LCMS (Method D): retention time 1.19 minutes, M / z = 584 (M + 1).

  5-Amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzo Diazole-5-carbonyl} piperidine-3-carbonitrile EV-AV4621-001 (EOAI3452077) I-82-step 8

  Tert-Butyl N- (5-cyano-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-yl in 10% trifluoroacetic acid (1 ml) with DCM Yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl) carbamate (EV-AV4615-004, 11 mg, 0.02 mmol) at room temperature. And allowed to stand for 1 hour. The reaction mixture was concentrated in vacuo and the residue was lyophilized from 1: 1 acetonitrile: water (4 ml) to give 5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo. [2,3-b] Pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-3-carbonitrile trifluoroacetic acid EV-AV4621-001 (I-82) [11.6 mg (quantitative)] was obtained as a white powder. LCMS (Method A): retention time 2.08 minutes, M / z = 484 (M + 1).

(1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-{[3- ( ² H ₃₎ methoxy-3-methyl cyclobutyl] methyl}-1H-1,3-benzo-oxadiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-amine EV-AY5000-002 (EOAI 3462946) I-238 is converted to methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy- as described in Scheme 3.8. 1-{[(1r, 3s) -3-hydroxy-3-ethylcyclobutyl] methyl} -1H-1,3-benzodiazole-5-carboxylate Scheme 3 by deuterium methylation of EV-AX8640-001 Obtained according to the method described in:

Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-{[(1r, 3s) -3- ( ² H ₃ ) Methoxy-3-methylcyclobutyl] methyl} -1H-1,3-benzodiazole-5-carboxylate EV-AX2096-001-Step 1

Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-{[(1r, 3s) -3-hydroxy-3-methyl To a stirred solution of cyclobutyl] methyl} -1H-1,3-benzodiazole-5-carboxylate (EV-AX8640-001, 85%, 250 mg, 0.45 mmol) / DMF (2.0 ml) was hydrogenated. Sodium (60%, 36 mg, 0.90 mmol) was added. The resulting mixture was stirred at room temperature for 10 minutes and iodo ( ² H ₃ ) methane (CAS865-50-9, 84 μl, 1.34 mmol) was added. The reaction mixture was stirred at room temperature for 4.5 hours and concentrated in vacuo. The residue was partitioned between EtOAc (20 ml) and water (15 ml). The aqueous layer was re-extracted with EtOAc (2 × 10 ml) and the combined organic layers were washed with water (10 ml), saturated aqueous sodium chloride solution (10 ml), dried over sodium sulfate and concentrated in vacuo. The resulting material was purified by flash chromatography (0-50% EtOAc / heptane) to give methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-{[(1r, 3s) -3- (2H-3-) methoxy-3-methylcyclobutyl] methyl} -1H-1,3-benzodiazole-5-carboxylate EV- AX2096-001 [184 mg (75%)] was obtained as a pale yellow solid. LCMS (Method D): retention time 1.43 minutes, M / z = 492 (M + 1).

  (1R, 4R, 7R) -2- {2- [6- (Difluoromethyl) -1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl- 1H-1,3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AW5368-001 (EOAI3460286) I-189 was described in Scheme 3.9. Methyl 2- [6- (difluoromethyl) -1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- Synthesized according to the method described in Scheme 3 via the synthesis of 5-carboxylate EV-AW5361-001:

  Methyl 2- {6-ethenyl-1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxy Rate EV-AW5353-002-Step 1

Methyl 2- {6-chloro-1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxy To a solution of rate (EV-AW5326-001,500 mg, 1.25 mmol) / dioxane (10 ml) was added tributyl (ethenyl) stannane (477 mg, 1.50 mmol). The reaction mixture was purged with nitrogen for 15 minutes, then Xantphos (54 mg, 0.04 mmol) and Pd ₂ dba ₃ (29 mg, 0.03 mmol) were added. The resulting mixture was stirred at 110 ° C. for 16 hours. The reaction mixture was concentrated in vacuo and the crude residue was purified by flash chromatography (0-50% EtOAc / heptane) to give methyl 2- {6-ethenyl-1-ethyl-1H-pyrrolo [2,3 -B] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AW5353-002 [340 mg (66%)] was obtained as a white solid. . LCMS (Method D): retention time 1.34 minutes, M / z = 391- (M + 1).

  Methyl 2- {1-ethyl-6-formyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxy Rate EV-AW5355-001-Step 2

Methyl 2- {6-ethenyl-1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxy To a solution of the rate (EV-AW5353-002, 89%, 470 mg, 1.07 mmol) / THF: water (2: 1, 9 ml), t-butanol (0.10 ml), 4-methylmorpholine-4-oxide ( 188 mg, 1.61 mmol) and OsO ₄ (0.10 ml / water, 0.02 mmol) were added. The reaction mixture was stirred at room temperature for 5 hours, NaIO ₄ (687 mg, 3.20 mmol) was added and stirring was continued at room temperature for 16 hours. The reaction mixture was filtered through a celite pad and washed with EtOAc. The filtrate was diluted with water (20 ml) and extracted with EtOAc (3 × 20 ml). The combined organic extracts were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (0-40% EtOAc / heptane) to give methyl 2- {1-ethyl-6-formyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-Methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AW5355-001 [270 mg (62%)] was obtained as a yellow solid. LCMS (Method D): retention time 1.24 minutes, M / z = 393 (M + 1).

  Methyl 2- [6- (difluoromethyl) -1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- 5-carboxylate EV-AW5361-001-step 3

  Methyl 2- {1-ethyl-6-formyl-1H-pyrrolo [2,3-b] pyridin-2-yl} -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxy To a solution of rate (200 mg, 0.50 mmol) / DCM (10 ml) was added 2-methoxy-N- (2-methoxyethyl) -N- (trifluoro-λ4-sulfanyl) ethanamine (CAS, 202289-38-1, 0.45 ml, 1.24 mmol) was added at room temperature. The reaction mixture was stirred at 45 ° C. for 16 hours, cooled to room temperature and poured into ice / water. The aqueous layer was neutralized with saturated sodium bicarbonate and extracted with DCM (3 × 20 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (0-40% EtOAc / heptane) to give methyl 2- [6- (difluoromethyl) -1-ethyl-1H-pyrrolo [2,3-b] pyridine-2. -Il] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AW5361-001 [105 mg (49%)] was obtained as a white solid. LCMS (Method D): retention time 1.33 minutes, M / z = 415 (M + 1).

  (1R, 4R, 7R) -2- {2- [6- (Difluoromethoxy) -1-ethyl-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl- 1H-1,3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AW9564-001 (EOAI 3460927) I-200 was described in Scheme 3.10. Synthesized according to the method described in Scheme 3 via synthesis of methyl 6- (difluoromethoxy) -1-ethyl-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AW9553-001:

Scheme 3.10

  2,2-Difluoro-2- (fluorosulfonyl) acetic acid (CAS 1717-59-5, 912 mg, 5.12 mmol) was added to benzyl N-[(3S, 4R) -1- {2- [1- (cyclopropylmethyl ) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -4-hydroxypiperidin-3-yl ] Was added to a stirred suspension of carbamate (EV-AW9553-001,451 mg, 2.05 mmol synthesized according to Scheme 3.18) and sodium sulfate (291 mg, 2.05 mmol) / acetonitrile (10 ml). The resulting mixture was stirred at room temperature for 16 hours. The crude product of the reaction was concentrated in vacuo and the residue was purified by flash chromatography (0-60% EtOAc / heptane) to give methyl 6- (difluoromethoxy) -1-ethyl-1H-pyrrolo [2 , 3-b] pyridine-2-carboxylate EV-AW9553-001 [327 mg (59%)] was obtained as an off-white solid. LCMS (Method D): retention time 1.30 minutes, M / z = 271 (M + 1).

  (1R, 4R, 7R) -2- {2- [7- (Cyclopropylmethyl) -2-methyl-7H-pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl -1H-1,3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AX1665-002 (EOAI 3460929) I-202 is described in Scheme 3.11. Methyl 2- [7- (cyclopropylmethyl) -2-methyl-7H-pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl-1H-1,3-benzodi Synthesized according to the method described in Scheme 3 via the synthesis of azole-5-carboxylate EV-AX 1646-002:

Methyl 2- [2-chloro-7- (cyclopropylmethyl) -7H-pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole 5-carboxylate (EV-AX1644-002,134mg, 0.31mmol) to a solution of / DME (5ml), potassium carbonate _{(87mg, 0.63mmol), PdCl 2} dppf (26mg, 0.03mmol) and Torimechiruboro Xone (3.5M in THF, 0.36ml, 1.26mmol) was added. The reaction mixture was stirred at 100 ° C. for 15 hours. The solvent is removed in vacuo and the crude residue is purified by flash chromatography (0-100% EtOAc / heptane then 0-40% methanol / EtOAc) to give methyl 2- [7- (cyclopropylmethyl) -2-Methyl-7H-pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AX1646-002 [ 94 mg (70%)] was obtained as an off-white solid. LCMS (Method D): retention time 1.05 min, M / z = 406 (M + 1).

  2- (2- {5-[(1R, 4R, 7R) -7-amino-2-azabicyclo [2.2.1] heptane-2-carbonyl] -7-methoxy-1-methyl-1H-1, 3-Benzodiazol-2-yl} -1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-6-yl) propan-2-ol EV-AW6283-001 (EOAI3461372) I- 210 is methyl 1- (cyclopropylmethyl) -6- (2-hydroxypropan-2-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV- as described in Scheme 3.12. Synthesized according to the method described in Scheme 3 by the synthesis of AW6273-002:

Scheme 3.12

  Methyl 6-acetyl-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AW6271-002-Step 1

Methyl 6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AW6269-001, 90%, 1.16 g, 3.93 mmol) / anhydrous dioxane (3 ml) was added to a solution of tributyl (1-ethoxyethenyl) stannane (1.59 ml, 4.71 mmol), Xantphos (0.17 g, 0.29 mmol) and Pd ₂ dba ₃ (0.09 g, 0.10 mmol). Was added. The reaction mixture was stirred at 90 ° C. for 17 hours. The solvent was removed in vacuo and 1M HCl (50 ml) and DCM (50 ml) were added to the residue. The biphasic mixture was stirred for 20 minutes, then the organic layer was separated and concentrated in vacuo. The crude residue was purified by flash chromatography (0-10% EtOAc / heptane) to give methyl 6-acetyl-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxyl. The rate EV-AW6271-002 [0.512 g (47%)] was obtained as an off-white solid. LCMS (Method D): retention time 1.40 minutes, M / z = 273 (M + 1).

Methyl 1- (cyclopropylmethyl) -6- (2-hydroxypropan-2-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AW6273-002 Step 2

  Methyl magnesium chloride (3M in THF, 642 μl) was added at −78 ° C. to methyl 6-acetyl-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV- AW6271-002, 510 mg, 1.84 mmol) / anhydrous THF (5 ml) was added dropwise. The reaction solution was stirred at −78 ° C. for 2.5 hours. Further methylmagnesium chloride (3M in THF, 61 μl) was kept stirring at −78 ° C. for 30 minutes. The reaction mixture was quenched with water (20 ml) and the THF was removed in vacuo. 1M HCl was added to the aqueous layer until pH 3 was reached. The aqueous layer was extracted with EtOAc (2 × 30 ml). The extracts were combined, dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (0-35% EtOAc / heptane) to give methyl 1- (cyclopropylmethyl) -6- (2-hydroxypropan-2-yl) -1H-pyrrolo [2, 3-b] pyridine-2-carboxylate EV-AW6273-002 [410 mg (76%)] was obtained as an off-white solid. LCMS (Method D): retention time 1.23 minutes, M / z = 289 (M + 1).

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfonylethyl) -7 -Methoxy-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AY0021-001 (EOAI 3461556) I-214 is prepared according to Scheme 3.13. 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfonylethyl) -7-methoxy-1H-1, Synthesized according to the method described in Scheme 3 via the synthesis of 3-benzodiazole-5-carboxylate EV-AY0016-001.

  (1R, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfinylethyl) -7 -Methoxy-1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AY0020-001 (EOAI 3461555) I-213 can be prepared according to Scheme 3.13. 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfinylethyl) -7-methoxy-1H-1, Synthesized according to the method described in Scheme 3 via the synthesis of 3-benzodiazole-5-carboxylate EV-AY0014-001.

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfinylethyl) -7-methoxy-1H-1,3-benzo Diazole-5-carboxylate EV-AY0014-001-Step 1

Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1- [2- (methylsulfanyl) ethyl] -1H-1,3 - benzodiazole 5-carboxylate (EV-AY0013-001,260mg, 0.52mmol) to a stirred solution of / methanol (8 ml) and water (2 ml), was added NaIO ₄ the (122mg, 0.57mmol). The reaction mixture was stirred at room temperature for 18 hours. The solvent was removed in vacuo and the residue was partitioned between EtOAc (20 ml) and water (50 ml). The aqueous layer was further extracted with EtOAc (2 × 20 ml). The organic layers were combined, washed with a saturated aqueous sodium chloride solution (20 ml), dried over sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography (0-50% EtOAc / heptane then 1-10% methanol / DCM) to give methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3- b] pyridin-2-yl] -1- (2-methanesulfinylethyl) -7-methoxy-1H-1,3-benzodiazole-5-carboxylate EV-AY0014-001 [220 mg (91%)] Obtained as a red foam. LCMS (Method D): retention time 1.08 min, M / z = 467 (M + 1).

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfonylethyl) -7-methoxy-1H-1,3-benzo Diazole-5-carboxylate EV-AY0016-001-Step 2

Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2-methanesulfinylethyl) -7-methoxy-1H-1,3-benzo diazole-5-carboxylate (EV-AY0014-001,135mg, 0.29mmol) to a stirred solution of / methanol (40 ml), was added KMnO ₄ a (50mg, 0.32mmol). The reaction mixture was stirred at room temperature for 15 minutes. The reaction was quenched by the addition of saturated aqueous sodium hydrogensulfate (20 ml). The mixture was filtered through Kieselguhr and the filtrate was concentrated in vacuo. The residue was diluted in EtOAc (20 ml) and water (50 ml) and the aqueous layer was extracted with EtOAc (2 × 20 ml). The organic layers were combined, washed with saturated aqueous sodium chloride solution (20 ml), dried over sodium sulfate, filtered and evaporated under reduced pressure to give methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [ 2,3-b] pyridin-2-yl] -1- (2-methanesulfonylethyl) -7-methoxy-1H-1,3-benzodiazole-5-carboxylate EV-AY0016-001 [115 mg (81 %)] As a brown solid. LCMS (Method D): retention time 1.15 minutes, M / z = 483 (M + 1).

  Rac-[(2R, 5S) -5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1- Methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-2-yl] methanol EV-AY4303-001 (EOAI3462115) I-220 and rac-[(2R, 5R) -5-amino-1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl } Piperidin-2-yl] methanol EV-AY4304-001 (EOAI3462116) I-221 is prepared according to the method described in Schemes 3 and 3.14 and rac-tert-butyl N-[(3R, 6S) -1- { 2- [1- (Siku Propylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (hydroxymethyl) Piperidin-3-yl] carbamate EV-AX4594-001 and rac-tert-butyl N-[(3R, 6R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b ] Pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4594-002 Obtained by the respective Boc-deprotection:

  Methyl 5-{[(tert-butoxy) carbonyl] amino} pyridine-2-carboxylate EV-AX4571-001-Step 1

  To a stirred solution of methyl 5-aminopyridine-2-carboxylate (CAS 67515-76-8, 1.8 g, 11.83 mmol) / DCM (40 ml) under a nitrogen atmosphere di-tert-butyl dicarbonate (2 .84 g, 13.01 mmol) and N, N-dimethylpyridin-4-amine (0.14 g, 1.18 mmol) were added and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was filtered (rinsing the filter with DCM) and the filtrate was diluted with DCM (100 ml) and washed with water (2 × 100 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give methyl 5-{[(tert-butoxy) carbonyl] amino} pyridine-2-carboxylate EV-AX4571-001 [2.73 g (60 %)] As an off-white solid. LCMS (Method D): retention time 1.01 min, M / z = 253 (M + 1).

  Tert-butyl N- [6- (hydroxymethyl) pyridin-3-yl] carbamate EV-AX4573-001-Step 2

  A stirred solution of methyl 5-{[(tert-butoxy) carbonyl] amino} pyridine-2-carboxylate (EV-AX4571-001, 2.73 g, 10.82 mmol) / THF (60 ml) at 0 ° C. with nitrogen Under atmosphere, 4M lithium tetrahydraluminate (1-) / diethyl ether (4.1 ml) was added dropwise and the reaction mixture was allowed to warm to room temperature and stirred for an additional 12 hours. The reaction was quenched with THF: water (9: 1, 15 ml) followed by 10% sodium hydroxide (10 ml) followed by water (10 ml). The reaction mixture is then filtered through a pad of Kieselguhr and rinsed with THF (3 × 50 ml). The filtrate is concentrated in vacuo and the crude product is purified by flash chromatography (50-100% EtOAc / heptane then 0-10% methanol / EtOAc) to give tert-butyl N- [6- (hydroxymethyl). ) Pyridin-3-yl] carbamate EV-AX4573-001 [1.50 g (60%)] was obtained as an off-white solid. LCMS (Method D): retention time 0.70 min, M / z = 224 (M + 1).

  Tert-Butyl acetate N- [6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4576-001-Step 3

In a pressure vessel, tert-butyl N- [6- (hydroxymethyl) pyridin-3-yl] carbamate (EV-AX4573-001, 1.50 g, 6.69 mmol) / ethanol (25 ml) and acetic acid (1.5 ml) A solution of was added. PtO ₂ (266 mg, 1.17 mmol) was added and the pressure vessel was purged with nitrogen, after which the reaction vessel was sealed and stirred at 65 ° C. for 16 hours under a hydrogen atmosphere (55 psi, 3.75 atm). . The reaction mixture was filtered through a pad of Kieselguhr, concentrated in vacuo and tert-butyl acetate N- [6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4576-001 [2.40 g (99%). Was obtained as an orange oil. No LCMS data.

  Benzyl 5-{[(tert-butoxy) carbonyl] amino} -2- (hydroxymethyl) piperidine-1-carboxylate EV-AX4584-001-Step 4

  Stirring tert-butyl acetate N- [6- (hydroxymethyl) piperidin-3-yl] carbamate (EV-AX4576-001, 70%, 2.00 g, 4.82 mmol) / THF (40 ml) and water (12 ml) To the solution was added sodium hydrogen carbonate salt (1.22 g, 14.47 mmol) and benzyl carbonochloridate (0.62 ml, 4.34 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C. for 15 minutes, the solution was allowed to warm to room temperature and stirring was continued for 4 hours. The reaction mixture was diluted with water (10 ml) and extracted with EtOAc (3 × 5 ml). The organic layers were combined, dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography (40-100% EtOAc / heptane) to yield benzyl 5-{[(tert-butoxy) carbonyl] amino} -2- (hydroxymethyl) piperidine-1-carboxylate EV. -AX4584-001 [1.40 g (80%)] was obtained as a white solid. LCMS (Method D): retention time 1.12 minutes, M / z = 387 (M + 23).

  Tert-butyl N- [6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4591-001-step 5

  To a solution of benzyl 5-{[(tert-butoxy) carbonyl] amino} -2- (hydroxymethyl) piperidine-1-carboxylate (EV-AX4584-001, 1.40 g, 3.84 mmol) / ethanol (20 ml) , Palladium on carbon (5%, 0.82 g, 0.38 mmol) was added and the reaction mixture was stirred for 14 hours at room temperature under hydrogen atmosphere. The mixture was filtered through a pad of Kieselguhr (washed with methanol) and the filtrate was concentrated in vacuo to give tert-butyl N- [6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4571- 001 [0.82 g (93%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.20 minutes, M / z = 231 (M + 1).

  Rac-tert-butyl N-[(3R, 6S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazol-5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4594-001 and rac-tert-butyl N-[(3R, 6R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- 5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4594-002-step 6

2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carboxylic acid To a stirred solution of (EV-AX2063-001, 200 mg, 0.53 mmol) / DMF (5 ml), DIPEA (463 μl, 2.66 mmol), HATU (222 mg, 0.58 mmol) and tert-butyl N- [6- ( Hydroxymethyl) piperidin-3-yl] carbamate (EV-AX4591-001, 122 mg, 0.53 mmol) was added at room temperature. The reaction mixture was stirred for 1 hour and concentrated in vacuo. The crude product was purified by preparative HPLC (acidification) to give two products:
rac-tert-butyl N-[(3R, 6S) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazol-5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4594-001 [70 mg (22%)] was obtained as an off-white solid. It was. LCMS (Method D): retention time 1.18 minutes, M / z = 589 (M + 1).

  rac-tert-butyl N-[(3R, 6R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazol-5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate EV-AX4594-002 [220 mg (70%)] was obtained as an off-white solid. It was. LCMS (Method D): retention time 1.19 minutes, M / z = 589 (M + 1).

  [(2S, 5R) -5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl- 1H-1,3-benzodiazol-5-carbonyl} piperidin-2-yl] methanol EV-AY4308-001 (EOAI 3462646) I-226 and [(2R, 5S) -5-amino-1- {2- [ 1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-2 -Yl] methanol EV-AY4309-001 (EOAI 3462647) I-227, both as described in Scheme 3, rac-tert-butyl N-[(3R, 6S) -1- {2- [1- ( Cyclopropylmethyl) -1H-pyrrolo [ 2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate EV Obtained by chiral resolution of -AX4594-001 followed by Boc-deprotection. The absolute configuration of both products was arbitrarily assigned.

  [(2R, 5R) -5-amino-1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl- 1H-1,3-benzodiazol-5-carbonyl} piperidin-2-yl] methanol EV-AY4310-001 (EOAI3462648) I-228 and [(2S, 5S) -5-amino-1- {2- [ 1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} piperidine-2 -Il] methanol EV-AY4311-001 (EOAI 3462649) I-229, both as described in Scheme 3, rac-tert-butyl N-[(3R, 6R) -1- {2- [1- ( Cyclopropylmethyl) -1H-pyro [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -6- (hydroxymethyl) piperidin-3-yl] carbamate Obtained by chiral resolution of EV-AX4594-002 followed by Boc-deprotection. The absolute configuration of both products was arbitrarily assigned.

  (1R, 4R, 7R) -2- {2- [7- (Cyclopropylmethyl) -2-methoxy-7H-pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl -1H-1,3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AX5554-001 (EOAI 346827) I-239 is described in Scheme 3.15 Synthesized according to the method described in Scheme 3 via the synthesis of methyl 7- (cyclopropylmethyl) -2-methoxy-7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AX5535-006 :

  Methyl 7- (cyclopropylmethyl) -2-methoxy-7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AX5535-006-Step 1

To a stirred solution of methyl 2-chloro-7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate (EV-AX2018-001, 1.00 g, 4.68 mmol) / anhydrous DMF (20 ml) at 0 ° C. Sodium hydride (60%, 0.28 g, 7.02 mmol) was added dropwise over 1 minute. (Bromomethyl) cyclopropane (680 μ, 7.01 mmol) was added at 0 ° C. for 30 minutes. The reaction mixture was warmed to room temperature and stirred for 60 hours. The reaction mixture was concentrated in vacuo and the resulting residue was partitioned between EtOAc (50 ml) and water (30 ml). The layers were separated and the organic phase was washed with 0.5M HCl (30 ml) and saturated aqueous sodium chloride solution (30 ml). The organic layer was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (5-40% EtOAc / heptane) to give 4 products:
Cyclopropylmethyl 2-chloro-7- (cyclopropylmethyl) -7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AX5535-003 [113 mg (8%)] as an off-white powder Obtained. LCMS (Method D): retention time 1.38 minutes, M / z = 306/308 (M + 1).
Methyl 2-chloro-7- (cyclopropylmethyl) -7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AX5535-004 [256 mg (20%)] was obtained as an off-white crystalline solid. . LCMS (Method D): 1.24 min, M / z = 266/268 (M + 1).
Cyclopropylmethyl 7- (cyclopropylmethyl) -2-methoxy-7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AX5535-005 [93 mg (6%)] was obtained as a yellow gum. It was. LCMS (Method D): retention time 1.32 min, M / z = 302 (M + 1).
Methyl 7- (cyclopropylmethyl) -2-methoxy-7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AX5535-006 [197 mg (16%)] was obtained as yellow crystals. LCMS (Method D): retention time 1.20 minutes, M / z = 262 (M + 1).

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AX4151-006 (EOAI34776815) I-269 is described in Scheme 3.16 1- (cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid was synthesized according to the method described in Scheme 3 via the synthesis of EV-AX4144-003:

Tert-butyl 6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AX4139-001-Step 1
6-Chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AU 7974-001, 500 mg, 1.99 mmol) / toluene (30 ml) with stirring The suspension was heated to 85 ° C. and N, N-dimethylformamide di-tert-butyl acetal (2.0 ml, 8.34 mmol) was added dropwise. The reaction mixture was stirred at 85 ° C. for 2 hours, and more N, N-dimethylformamide di-tert-butyl acetal (2.0 ml, 8.34 mmol) was added. Stirring was continued at 85 ° C. for 18 hours and then at 100 ° C. for 3 hours. After cooling, the reaction mixture was diluted with EtOAc (200 ml), washed with saturated aqueous sodium chloride solution (50 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography (0-10% EtOAc / heptane) to give tert-butyl 6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2. -Carboxylate EV-AX4139-005 [366 mg (60%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.66 minutes, M / z = 307/309 (M + 1).

  Tert-butyl 1- (cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AX4142-001-step 2

  tert-Butyl 6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AX4139-005, 366 mg, 1.19 mmol) / DMF (5 ml) To the stirring solution was added tetramethylammonium fluoride (CAS 373-68-2, 250 mg, 2.68 mmol) and the mixture was stirred at 80 ° C. for 18 hours. After cooling, the reaction mixture was partitioned between DCM (200 ml), water (100 ml) and saturated aqueous sodium chloride solution (100 ml). The organic layer was separated and further washed with saturated aqueous sodium chloride solution (50 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude material was purified by flash chromatography (0-10% EtOAc / heptane) to give tert-butyl 1- (cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridine-2- The carboxylate EV-AX4142-001 [112 mg (32%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.61 min, M / z = 291- (M + 1).

  1- (cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AX4144-003-step 3

  tert-Butyl 1- (cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AX4142-001, 100 mg, 0.34 mmol) / TFA (2.0 ml) , 26.2 mmol) was stirred at room temperature for 3 hours. After concentration in vacuo, the residue was taken up in DCM and concentrated to 1- (cyclopropylmethyl) -6-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AX4144-003. [80 mg (89%)] was obtained as a yellow solid. LCMS (Method D): retention time 1.15 minutes, M / z = 235 (M + 1).

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -6- (oxan-4-yl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7 -Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AY7194-001- (EOAI34707013) I-275 Synthesis of ethyl 1- (cyclopropylmethyl) -6- (oxan-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AY7184-001 as described in Scheme 3.17 Was synthesized according to the method described in Scheme 3:

  Ethyl 1- (cyclopropylmethyl) -6- (3,6-dihydro-2H-pyran-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AY7182-001-step 1

Ethyl stirred solution 6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AY7177-001, 90%, 950 mg, 3.07 mmol) and 2- (3,6-dihydro-2H-pyran-4-yl) -4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS287944-16-5,709 mg, 3.37 mmol) was added to THF. : Dissolved in toluene (4: 1, 10 ml) and 2M sodium carbonate (3.07 ml) was added. The reaction mixture was purged with nitrogen for 5 minutes and Pd (dppf) Cl ₂ (449 mg, 0.61 mmol) was added. The reaction mixture was stirred at 100 ° C. for 4 hours, cooled to room temperature and concentrated in vacuo. The resulting residue was dissolved in EtOAc (20 ml) and washed with water (2 × 10 ml) and saturated aqueous sodium chloride solution (10 ml). The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash chromatography (0-50% EtOAc / heptane) to give ethyl 1- (cyclopropylmethyl) -6- (3,6-dihydro-2H-pyran-4-yl) -1H- Pyrrolo [2,3-b] pyridine-2-carboxylate EV-AY7182-001 [587 mg (56%)] was obtained as an orange oil. LCMS (Method D): 1.52 min, M / z = 327 (M + 1).

  Ethyl 1- (cyclopropylmethyl) -6- (oxan-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AY7184-001-Step 2

  Ethyl 1- (cyclopropylmethyl) -6- (3,6-dihydro-2H-pyran-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AY7182-001) To a stirred solution of 587 mg, 1.71 mmol) / EtOAc: ethanol (1: 1, 30 ml) was added Pd / C (10%, 91 mg, 0.09 mmol). The reaction mixture was left under a hydrogen atmosphere and stirred at room temperature for 18 hours. The reaction mixture was filtered through a Kieselguhr pad and washed with methanol (30 ml). The filtrate was concentrated in vacuo to give ethyl 1- (cyclopropylmethyl) -6- (oxan-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AY7184-001 [ 516 mg (83%)] was obtained as an off-white powder. LCMS (Method D): retention time 1.51 min, M / z = 329 (M + 1).

(1R, 4R, 7R) -2- {2- [1- ( cyclopropylmethyl) -6- ⁽² H ₃₎ methoxy -1H- pyrrolo [2,3-b] pyridin-2-yl] -1 Methyl-1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AZ5120-001- (EOAI3478073) I-285 is shown in Scheme 3.18. through the synthesis of methyl describes 1- (cyclopropylmethyl) -6- ⁽² H ₃₎ methoxy -1H- pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ5103-001 in scheme 3 Synthesized according to the method described in:

  1- (Cyclopropylmethyl) -6-hydroxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AX5573-002-Step 1

Ethyl 6-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AY7167-001, 90%, 1.45 g, 4.68 mmol), Pd ₂ (dba) ₃ (214 mg, 0.23 mmol), ^t Bu-BippyPhos (237 mg, 0.47 mmol) and potassium hydroxide (788 mg, 14.0 mmol) in a dioxane (7 ml) and water (7 ml) in a pressure tube. Combined. The reaction mixture was purged with nitrogen for 5 minutes, then the vessel was sealed and heated at 70 ° C. for 1.5 hours. The reaction mixture was cooled to room temperature and filtered through glass fiber filter paper. The filtrate was partitioned between water (10 ml) and EtOAc (30 ml). The aqueous layer is acidified to pH 5 using 2M HCl and the resulting precipitate is filtered to give 1- (cyclopropylmethyl) -6-hydroxy-1H-pyrrolo [2,3-b] pyridine-2- The carboxylic acid EV-AX5573-002 [0.80 g (62%)] was obtained as an off-white powder. LCMS (Method D): retention time 0.95 min, M / z = 233 (M + 1).

  Methyl 1- (cyclopropylmethyl) -6-hydroxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AY4950-002-Step 2

  1- (Cyclopropylmethyl) -6-hydroxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AX5573-002, 73%, 550 mg, 1.73 mmol) / anhydrous toluene (6 ml) And to a stirred suspension of anhydrous methanol (2 ml, 49.44 mmol) was added 2M (diazomethyl) (trimethyl) silane (1.73 ml in diethyl ether) under a nitrogen atmosphere. The resulting mixture was stirred for 1.5 hours at room temperature. Acetic acid (˜0.7 ml) was added until the light yellow color disappeared. The reaction mixture was concentrated in vacuo and triturated with DCM (5 ml). The solid was filtered off and dried to give 1- (cyclopropylmethyl) -6-hydroxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid EV-AY4950-001 [131 mg (28%) ] Was obtained as a light skin-colored solid. LCMS (Method D): retention time 1.13 minutes, M / z = 247 (M + 1).

Methyl 1- (cyclopropylmethyl) -6- ⁽² H ₃₎ methoxy -1H- pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ5013-001- Step 3

To a stirred solution of 1- (cyclopropylmethyl) -6-hydroxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AY4950-001, 100 mg, 0.40 mmol) / DMF (30 ml) , Sodium hydride (60%, 32 mg, 0.80 mmol) was added. The reaction mixture was stirred at room temperature for 10 minutes and then iodo ( ² H ₃ ) methane (74 μl, 1.19 mmol) was added. The reaction mixture was stirred at room temperature for 17 hours, diluted with EtOAc (20 ml) and washed with water (2 × 20 ml) and saturated aqueous sodium chloride solution (10 ml). The organic phase was dried, concentrated in vacuo, methyl 1- (cyclopropylmethyl) -6- ⁽² H ₃₎ methoxy -1H- pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ5013- 001- (85 mg, 75%) was obtained as a yellow powder. LCMS (Method D): retention time 1.36 minutes, M / z = 264 (M + 1).

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -3-fluoro-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AZ5131-001- (EOAI3478196) I-292 in scheme 3.19 Synthesis according to the method described in Scheme 3 via the synthesis of the described ethyl 1- (cyclopropylmethyl) -3-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ5111-001 did:

  Ethyl 1- (cyclopropylmethyl) -3-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ5111-001-Step 1

  To a stirred solution of ethyl 1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AZ5106-001, 1.00 g, 4.09 mmol) / DMF (20 ml). , Selectfluor® (CAS 140681-55-6, 1.45 g, 4.09 mmol) was added. The resulting mixture was stirred at room temperature for 60 hours. The reaction mixture is concentrated in vacuo and the residue is dissolved in EtOAc (20 ml) and washed with water (3 × 20 ml). The organic phase was dried over sodium sulfate and concentrated in vacuo. The crude product was purified by flash chromatography (0-50% DCM / heptane) to give ethyl 1- (cyclopropylmethyl) -3-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxy. The rate EV-AZ5111-001 [245 mg (22%)] was obtained as a yellow oil. LCMS (Method D): retention time 1.33 minutes, M / z = 263 (M + 1).

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -4-fluoro-1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl -1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AY4588-001 (EOAI3478689) I-294 is shown in Schemes 3.16 and 3 Through the synthesis of tert-butyl 1- (cyclopropylmethyl) -4-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AX4168-003 as described in .20 Synthesized according to the method described:

  Tert-butyl 1- (cyclopropylmethyl) -4-fluoro-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AX4168-003-step 1

  tert-Butyl 4-chloro-1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AX4167-001, 820 mg, 2.67 mmol) / anhydrous DMF (10 ml) To the stirred solution was added tetramethylammonium fluoride (500 mg, 5.36 mmol) and the mixture was stirred at 80 ° C. for 18 hours. The reaction mixture was partitioned between DCM (200 ml) and saturated aqueous sodium bicarbonate (200 ml). The aqueous layer was washed with DCM (2 × 100 ml) and the combined organic layers were washed with saturated aqueous sodium chloride solution (100 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude material was purified by flash chromatography (0-10% EtOAc / heptane) to give tert-butyl 1- (cyclopropylmethyl) -4-fluoro-1H-pyrrolo [2,3-b] pyridine-2- The carboxylate EV-AX4168-003 [401 mg (52%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.48 minutes, M / z = 291- (M + 1).

  (1R, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -6- (morpholin-4-yl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7 -Methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine EV-AX5592-001 (EOAI3478190) I-287, Synthesis of ethyl 1- (cyclopropylmethyl) -6- (morpholin-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ2525-001 described in Scheme 3.21. Via the method described in Scheme 3:

  Ethyl 1- (cyclopropylmethyl) -6- (morpholin-4-yl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AZ2525-001-Step 1

Morpholine (74 μl, 0.86 mmol), Pd ₂ (dba) ₃ (33 mg, 0.04 mmol), Xantphos (41 mg, 0.07 mmol) and sodium-tert-butoxide (103 mg, 1.07 mmol) were combined with ethyl 6-bromo. A nitrogen purged solution of -1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AZ2517-001, 231 mg, 0.71 mmol) / toluene (6 ml) was added. Added to the pressure tube. The pressure tube was sealed under a nitrogen environment. The reaction mixture was heated at 110 ° C. for 2 hours, cooled to room temperature, diluted with water (10 ml) and extracted with EtOAc (3 × 10 ml). The combined organic layers were washed with saturated aqueous sodium chloride solution (10 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography (0-20% EtOAc / heptane) to give ethyl 1- (cyclopropylmethyl) -6- (morpholin-4-yl) -1H-pyrrolo [2,3-b. ] Pyridine-2-carboxylate EV-AZ2525-001 [60 mg (25%)] was obtained as a yellow solid. LCMS (Method D): retention time 1.35 minutes, M / z = 330 (M + 1).

5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine -2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole hydrochloride EV-AV3043-001 (EOAI3452073, absolute stereochemistry arbitrarily assigned) of I-24 Composition

  Methyl 4-chloro-3-hydroxy-5-nitrobenzoate EV-AU3635-002-step 1

  Note: This reaction was performed twice under the same conditions. The crude from each reaction was combined after quenching with methanol as described below.

To a stirred solution of methyl 4-chloro-3-methoxy-5-nitrobenzoate (CAS 66033-09-8, 5.00 g, 20.4 mmol) / anhydrous DCM (20 ml) at 0 ° C. under nitrogen under BBr ₃ (1M in DCM, 40.71 ml, 2.81 mmol) was added dropwise over 20 minutes. The reaction mixture was stirred at 0 ° C. for 30 minutes, then allowed to warm to room temperature and stirred for 15 hours. The reaction mixture was cooled to 0 ° C., carefully quenched with methanol, combined and concentrated in vacuo. The residue was dissolved in methanol (300 ml), concentrated, sulfuric acid (10 drops) was added and the reaction mixture was stirred at 75 ° C. for 5 h. The cooled mixture was concentrated in vacuo and to the residue was carefully added water (50 ml) and saturated NaHCO ₃ (50 mL) to reach a basic pH. The suspension was sonicated for 15 minutes and stirred for an additional 30 minutes, after which the resulting solid was collected and dried under vacuum filtration. The solid was washed with water (25 ml) and dried to give methyl 4-chloro-3-hydroxy-5-nitrobenzoate EV-AU3635-002 [8.14 g (82.9%)] as a light brown powder. LCMS (Method D): retention time 1.11 min, M / z = 230 (M + 1).

  Methyl 4-chloro-3- (difluoromethoxy) -5-nitrobenzoate EV-AT8693-001-step 2

  Note: This reaction was performed three times under the same conditions. The crude from each reaction was collected for workup as described below.

Methyl 4-chloro-3-hydroxy-5-nitrobenzoate (EV-AU3635-002, 333 mg, 4.32 mmol) and potassium carbonate (7.17 g, 155.4 mmol) / acetonitrile: water (1: 1, 20 ml) solution was added 2-chloro-2,2-difluoro-1-phenylethanone (1.06 ml, 21.6 mmol). The vessel was sealed and heated at 80 ° C. for 16 hours. The cooled reaction mixtures were combined and partitioned between EtOAc (200 ml) and 2M HCl (aq). The aqueous fraction (at pH 5) was back extracted with more EtOAc (2 × 100 ml). The organic layers were combined, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was dissolved in methanol (150 ml), concentrated sulfuric acid (3 drops) was added and the reaction mixture was stirred at 75 ° C. for 40 hours. The cooled mixture was concentrated in vacuo and saturated NaHCO ₃ was carefully added to the residue until a basic pH was reached. The aqueous layer was extracted with EtOAc (3 × 100 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by flash chromatography (0-20% EtOAc / heptane) to give methyl 4-chloro-3- (difluoromethoxy) -5-nitrobenzoate EV-AT8693-001 [775 mg (60%)] Was obtained as a yellow solid. LCMS (Method D): retention time 1.20 minutes, no mass ions were observed.

  Methyl 3- (difluoromethoxy) -4- (methylamino) -5-nitrobenzoate EV-AV3021-001-step 3

Methyl 4-chloro-3- (difluoromethoxy) -5-nitrobenzoate (EV-AT8693-001,400mg, 1.34mmol) / To a stirred solution of _{THF (10ml), K 2 CO} 3 (1.29g, 9. 35 mmol) and methanamine (2.0 M in THF, 1 ml, 2.00 mmol) were added. The reaction mixture was stirred for 24 hours at room temperature, concentrated in vacuo and partitioned between EtOAc (30 ml) and 1M HCl (15 ml). The organic fraction was washed with additional 1M HCl (15 ml), saturated aqueous sodium chloride solution (10 ml), dried over sodium sulfate, filtered, concentrated in vacuo and methyl 3- (difluoromethoxy) -4- ( Methylamino) -5-nitrobenzoate EV-AV3021-001 (370 mg, 88%) was obtained as an orange powder. LCMS (Method D): retention time 1.17 minutes, M / z = 277 (M + 1).

  Methyl 3-amino-5- (difluoromethoxy) -4- (methylamino) benzoate EV-AV3024-001-step 4

  To a stirred solution of methyl 3- (difluoromethoxy) -4- (methylamino) -5-nitrobenzoate (EV-AV3021-001, 370 mg, 1.18 mmol) / EtOAc: EtOH (1:20 ml) under nitrogen 10% Pd / C (62.7 mg, 0.06 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours under hydrogen atmosphere. The reaction mixture was filtered through Kieselguhr and the filter cake was washed with EtOAc. The filtrate was concentrated in vacuo to give methyl 3-amino-5- (difluoromethoxy) -4- (methylamino) benzoate EV-AV3024-001 [296 mg (93%)] as a light brown powder. LCMS (Method D): retention time 0.97 min, M / z = 247 (M + 1).

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole 5-carboxylate EV-AV3026-001-step 5

  1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AS5740-003, 139 mg, 0.55 mmol synthesized according to Scheme 1, Step 1) / DMF (5 ml) To a stirred solution of was added DIPEA (0.08 ml, 0.47 mmol) followed by HATU (177 mg, 0.47 mmol) at 0 ° C. The resulting mixture was stirred at 0 ° C. for 10 minutes, methyl 3-amino-5- (difluoromethoxy) -4- (methylamino) benzoate (EV-AV3024-001, 150 mg, 0.55 mmol) was added, The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was then stirred at 50 ° C. for 22 hours and then concentrated in vacuo. The crude residue was dissolved in acetic acid (3 ml) and heated in a sealed tube at 80 ° C. for 2 hours. The solvent was removed in vacuo and the residual material was purified by flash chromatography (0-70% EtOAc / heptane) to give methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b. ] Pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5-carboxylate EV-AV3026-001 [131 mg (48%)] was pale brownish As a white powder. LCMS (Method D): retention time 1.29 minutes, M / z = 427 (M + 1).

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5 -Carboxylic acid EV-AV3032-001-Step 6

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole To a stirred solution of 5-carboxylate (EV-AV3026-001, 131 mg, 0.26 mmol) / THF: MeOH (4 ml: 1 ml) was added 2M lithium hydroxide (528 μl, 1.06 mmol) and the reaction mixture was added to 40 mL. Stir at 16 ° C. for 16 hours. The reaction mixture was concentrated in vacuo, suspended in water (2 ml) and acidified to pH 2 using 2M HCl. The resulting suspension was stirred for 10 minutes and the resulting precipitate was collected by vacuum filtration and dried to give 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b ] Pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5-carboxylic acid EV-AV3032-001 [108 mg (97%)] as an orange powder Obtained. LCMS (Method D): retention time 1.15 minutes, M / z = 413 (M + 1).

  Tert-butyl 6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3 -Benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AV3033-001-Step 7

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5 A solution of carboxylic acid (EV-AV3032-001, 108 mg, 0.26 mmol) / DMSO (2 ml) at 0 ° C. with HATU (117.1 mg, 0.31 mmol) and DIPEA (87.87 μl, 0.51 mmol) Was added. The reaction mixture was stirred for 10 minutes, tert-butyl octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (CAS 949559-11-9, 61 mg, 0.27 mmol) was added and the reaction mixture was added. Was stirred at room temperature for 16 hours. The reaction mixture was diluted with water (1 ml) and purified by preparative HPLC (basic method) to give tert-butyl 6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2, 3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine -1-Carboxylate EV-AV3033-001 [125 mg (79%)] was obtained as a white powder. LCMS (Method A): retention time 3.96 minutes, M / z = 621- (M + 1).

  tert-Butyl (3aS, 7aR) -6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl -1H-1,3-benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AV3033-002 and tert-butyl (3aR, 7aS) -6 -{2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole Chiral HPLC to obtain -5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AV3033-003—Step 8

  tert-butyl 6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3 -Benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AV3033-001 [125 mg] was dissolved in methanol and then chiral HPLC (Method G ) To give tert-butyl (3aS, 7aR) -6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoro Methoxy) -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AV3033-002 [47.6 mg ( 27%)] And tert-butyl (3aS, 7aS) -6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1- Methyl-1H-1,3-benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate EV-AV3033-003 [50.1 mg (29%)] Both were obtained as colorless gums (the absolute stereochemistry was arbitrarily assigned).

EV-AV3033-002 chiral purity (UV, 254 nm): 100%, retention time: 5.46 minutes (Method I)
EV-AV3033-003 chiral purity (UV, 254 nm): 100%, retention time: 7.87 min (Method I)

  5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine -2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole hydrochloride EV-AV3043-001 (EOAI3452073, absolute stereochemistry arbitrarily assigned) I-24 Step 9

  Tert-butyl (3aS, 7aR) -6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl -1H-1,3-benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AV3033-002, 47.6 mg, 0.07 mmol) Dissolved in 1.25 M HCl / EtOH (1 ml) and stirred at 40 ° C. for 6 hours. The reaction mixture was concentrated in vacuo, dissolved in water (2 ml) and lyophilized to give 5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl]. 2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole hydrochloride Salt EV-AV3043-001 (I-24) [29 mg (71%)] was obtained as a yellow powder. LCMS (Method A): retention time 2.23 minutes, M / z = 521- (M + 1).

  5-[(3aR, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine -2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole hydrochloride EV-AV3044-001 (EOAI 3345704, absolute stereochemistry assigned arbitrarily) I-25 Step 9

  Tert-butyl (3aS, 7aS) -6- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl -1H-1,3-benzodiazole-5-carbonyl} -octahydro-1H-pyrrolo [2,3-c] pyridine-1-carboxylate (EV-AV3033-003, 50.1 mg, 0.08 mmol) Dissolved in 1.25 M HCl / EtOH (1 ml) and stirred at 40 ° C. for 6 hours. The reaction mixture was concentrated in vacuo, dissolved in water (2 ml) and lyophilized to give 5-[(3aR, 7aS) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl]. 2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole hydrochloride Salt EV-AV3044-001 (I-25) [37 mg (87%)] was obtained as a yellow powder. LCMS (Method A): retention time 2.26 minutes, M / z = 521- (M + 1).

  Methyl 2-methoxy-4- (methylamino) -5-nitrobenzoate EV-AS3796-002-Step 1

  To a solution of methyl 4-chloro-2-methoxy-5-nitrobenzoate (CAS 10909069-75-2, 1.50 g, 6.11 mmol) / DMF (15 ml) at 0 ° C., methylamine (2.0 M in THF) was added. , 3.66 ml, 7.33 mmol) was added and the resulting reaction mixture was allowed to warm to room temperature and stirred for 20 hours. Potassium carbonate (1.01 g, 7.33 mmol) was added and the reaction mixture was stirred at room temperature for 1 h and additional methylamine (2.0 M in THF, 3.66 ml, 7.33 mmol) was added. . The reaction mixture was stirred for an additional 5 hours at room temperature. The reaction mixture is concentrated in vacuo and the crude residue is purified using flash column chromatography (0-100% EtOAc / heptane then 50% EtOAc / methanol) to give methyl 2-methoxy-4- (methyl Amino) -5-nitrobenzoate EV-AS3796-002 [905 mg (56%)] was obtained as a yellow powder. LCMS (Method D): retention time 1.04 minutes, M / z = 241- (M + 1).

  Methyl 5-amino-2-methoxy-4- (methylamino) benzoate EV-AS3799-003 Step 2

  To a stirred solution of methyl 2-methoxy-4- (methylamino) -5-nitrobenzoate (EV-AS3796-002, 905 mg, 3.77 mmol) / ethanol: EtOAc (1: 2, 60 ml) under nitrogen. Pd / C (10%, 200 mg, 0.19 mmol) was added and the reaction mixture was stirred under a hydrogen atmosphere for 5.5 hours. The reaction mixture was filtered through Kieselguhr and the filter cake was washed through with EtOAc, ethanol and DCM. The filtrate was concentrated in vacuo and the residue was dissolved in DCM: ethanol (6: 1, 70 ml). Pd / C (10%, 200 mg, 0.19 mmol) was added under nitrogen and the reaction mixture was stirred under a hydrogen atmosphere for 6.5 hours. The reaction mixture was filtered through Kieselguhr and the filter cake was washed through with ethanol and DCM. The filtrate was concentrated in vacuo and the residue was dissolved in DCM: ethanol (1: 1, 120 mL). Pd / C (10%, 200 mg, 0.19 mmol) was added under nitrogen and the reaction mixture was stirred under a hydrogen atmosphere for 22 hours. The reaction mixture was filtered through Kieselguhr and the filter cake was washed through with ethanol and DCM. The filtrate was concentrated in vacuo to give methyl 5-amino-2-methoxy-4- (methylamino) benzoate EV-AS3799-003 [656 mg (76%)] as a dark brown powder. LCMS (Method A): retention time 0.90 min, M / z = 211 (M + 1).

  According to the method described in Scheme 1 using EV-AS3799-003, (3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2- IL] -6-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-amine hydrochloride EV-AV3612-001 (EOAI 3447040) I-68 was synthesized.

  Methyl 2-methyl-4- (methylamino) -5-nitrobenzoate EV-AV4643-001-step 1

To a stirred solution of methyl 4-fluoro-2-methyl-5-nitrobenzoate (CAS1163287-01-1, 1.00 g, 4.69 mmol) / THF (12 ml) was added methylamine (2.0 M in THF, 5. 4 ml, 10.8 mmol) was added and the reaction mixture was stirred at room temperature for 10 minutes. The reaction mixture was partitioned between ethyl acetate (250 ml) and saturated aqueous NaHCO ₃ (100 ml). The organic extract was washed with water (100 ml), dried over sodium sulfate, filtered, concentrated in vacuo and methyl 2-methyl-4- (methylamino) -5-nitrobenzoate EV-AV4643-001 [1 0.07 g (quantitative)] was obtained as a yellow powder. LCMS (Method D): retention time 1.14 minutes, M / z = 225 (M + 1).

  Methyl 5-amino-2-methyl-4- (methylamino) benzoate EV-AV4644-001-Step 2

  To a stirred solution of methyl 2-methyl-4- (methylamino) -5-nitrobenzoate (EV-AV4643-001, 1.07 g, 4.77 mmol) / ethanol (100 ml) was added 10% Pd / C under nitrogen. (102 mg, 0.048 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction mixture was filtered through glass fiber sinter and the filtrate was concentrated in vacuo to give methyl 5-amino-2-methyl-4- (methylamino) benzoate EV-AV4644-001 [1.02 g (98%)]. , Obtained as a light pale brown crystalline solid. LCMS (Method D): retention time 0.77 min, M / z = 195 (M + 1).

  Methyl 5-amino-2-methyl-4- (methylamino) benzoate EV-AV4644-001 according to the method described in Scheme 2 (3R) -1- {2- [1- (cyclopropylmethyl ) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1,6-dimethyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-amine hydrochloride EV-AV4649- 001 (EOAI 3454825) I-103 was synthesized.

5-[(3R) -3-Aminopiperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H Of 1,3-benzodiazole-7-carboxamide hydrochloride EV-AV9635-001 (EOAI3455108) I-112

  Phenyl 5-[(3R) -3-{[(tert-butoxy) carbonyl] amino} piperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine -2-yl] -1-methyl-1H-1,3-benzodiazole-7-carboxylate EV-AV9626-002-Step 1

A solution of Pd (OAc) ₂ (6 mg, 0.03 mmol) and Xantphos (30 mg, 0.05 mmol) in acetonitrile (20 ml) in a pressure tube was degassed for 5 minutes to give tert-butyl N-[(3R). -1- {7-bromo-2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole- 5-carbonyl} piperidin-3-yl] carbamate (EV-AV9622-001 synthesized according to Scheme 1 (520 mg, 0.86 mmol), phenyl formate (0.19 ml, 1.71 mmol) and triethylamine (0.24 ml, 1. The reaction vessel was sealed and heated for 4 hours at 80 ° C. The cooled reaction mixture was diluted with EtOAc and washed with water. The crude residue was purified by flash chromatography (0-100% EtOAc) to give phenyl 5-[(3R) -3-{[(tert-butoxy ) Carbonyl] amino} piperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3- Benzodiazole-7-carboxylate EV-AV9626-002 [220 mg (39%)] was obtained as a flesh-colored solid LCMS (Method D): retention time 1.35 minutes, M / z = 649 (M + 1).

  Tert-butyl N-[(3R) -1- {7-carbamoyl-2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9632-002-Step 2

  Phenyl 5-[(3R) -3-{[(tert-butoxy) carbonyl] amino} piperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3- b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazole-7-carboxylate (EV-AV9626-002, 110 mg, 0.17 mmol) / dioxane (5 ml) in water Ammonium oxide (35% w / w, 0.1 ml, 2.54 mmol) was added. The vessel was sealed and heated at 50 ° C. for 2.5 hours. Additional ammonium hydroxide (35% w / w, 0.5 ml) was added and the vessel was sealed and heated at 50 ° C. for 17 hours. Additional ammonium hydroxide (35% w / w, 0.5 ml) was added and the vessel was sealed and stirred at 50 ° C. for a further 23 hours. The reaction mixture was concentrated in vacuo, redissolved in DCM and washed with 5% NaOH (aq). The organic fraction was passed through a phase separation cartridge and concentrated in vacuo. The crude residue was purified by preparative HPLC (basic method) and tert-butyl N-[(3R) -1- {7-carbamoyl-2- [1- (cyclopropylmethyl) -1H-pyrrolo]. [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate EV-AV9632-002 [75 mg (77%) Was obtained as an off-white powder. LCMS (Method D): retention time 1.08 min, M / z = 572 (M + 1).

  5-[(3R) -3-Aminopiperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazole-7-carboxamide hydrochloride EV-AV9635-001- (I-112) -step 3

  tert-Butyl N-[(3R) -1- {7-carbamoyl-2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-yl] carbamate (EV-AV9632-002, 75 mg, 0.13 mmol) was added 1.25 M HCl / ethanol (3 ml) and the reaction The mixture was stirred at 40 ° C. for 1.5 hours. The reaction mixture was concentrated in vacuo and the residue was lyophilized from water (3 mL) to give 5-[(3R) -3-aminopiperidine-1-carbonyl] -2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzodiazol-7-carboxamide hydrochloride EV-AV9635-001 (I-112) [57. 6 mg (84%)] was obtained as an off-white powder. LCMS (Method A): retention time 1.53 minutes, M / z = 472 (M + 1).

  Special case

  2- [7- (Cyclopropylmethyl) -7H-pyrrolo [2,3-d] pyrimidin-6-yl] -1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6 -Carbonyl} -1H-1,3-benzodiazole EV-AQ4191-002 (EOAI3434971) I-9 was synthesized according to the method described in Scheme 1 to give 2- [7- (cyclopropylmethyl) -7H- Pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl-5- {octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl} -1H-1,3 -Benzodiazole EV-AS1566-001 (EOAI3435740) I-15, 5-[(3aR, 7aR) -octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- [7- ( Cyclopropylmethyl) -7H Pyrrolo [2,3-d] pyrimidin-6-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole EV-AS1590-001 (EOAI3437977) I-12 and 5-[(3aS, 7aS) -Octahydro-1H-pyrrolo [2,3-c] pyridine-6-carbonyl] -2- [7- (cyclopropylmethyl) -7H-pyrrolo [2,3-d] pyrimidin-6-yl]- 7-methoxy-1-methyl-1H-1,3-benzodiazole EV-AS1591-001 (EOAI 3437978) I-4 was converted to methyl 7H-pyrrolo [2,3-d] pyrimidine-6- 6 described in Scheme 7. Synthesized according to the method described in Scheme 2 via the synthesis of carboxylate EV-AQ 1970-001:

  To a stirred suspension of 7H-pyrrolo [2,3-d] pyrimidine-6-carboxylic acid (CAS1016241-64-7, 0.50 g, 3.06 mmol) / methanol (10 ml) at 0 ° C. under a nitrogen atmosphere Below, thionyl dichloride (0.56 ml, 7.66 mmol) was added dropwise. The resulting mixture was allowed to warm to room temperature and then heated to reflux for 24 hours. Additional methanol (15 ml) was added to aid dissolution, the mixture was cooled to 0 ° C. and more thionyl dichloride (0.56 ml, 7.66 mmol) was added dropwise under a nitrogen atmosphere. The mixture was heated at reflux for 3 hours, evaporated to dryness and methyl 7H-pyrrolo [2,3-d] pyrimidine-6-carboxylate EV-AQ 1970-001 [0.66 g (quantitative)] Obtained as a colored powder. LCMS (Method J): retention time 1.02 min, M / z = 178 (M + 1).

  (3R) -1- {2- [1- (Cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl] -1-methyl-1H-1,3-benzo Diazole-5-carbonyl} piperidin-3-amine EV-AU3631-001 (EOAI34447868) I-70 was synthesized according to the method described in Scheme 1 to give (3R) -1- {2- [1- (cyclo Propylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl] -1- (2,2-difluoroethyl) -7-methoxy-1H-1,3-benzodiazole- 5-carbonyl} piperidin-3-amine EV-AV3086-001 (EOAI3454400) I-94 and (3R) -1- {2- [1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3 -B] pyridin-2-yl] -7- Toxi-1-methyl-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-amine EV-AV4830-003 (EOAI3454405) I-33 was synthesized according to the method described in Scheme 2, 3R) -1- {2- [1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H -1,3-benzodiazol-5-carbonyl} piperidin-3-amine EV-AV3852-001 (EOAI 3454816) I-104 and (3R, 5R) -1- {2- [1- (cyclopropylmethyl)- 6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl] -7- (difluoromethoxy) -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -5-fluor Piperidine-3-amine EV-AV4849-001 (EOAI3455105) I-109 was converted to ethyl 6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AU3621-002 described in Scheme 8. Via synthesis, synthesized according to the method described in Scheme 3:

  1- (Benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine EV-AT6244-001-Step 1

  To a stirred solution of 6-methoxy-1H-pyrrolo [2,3-b] pyridine (CAS896722-53-5,300 mg, 2.02 mmol) / THF (15 ml) at 0 ° C., sodium hydride (60%, 121 mg , 3.04 mmol) was added dropwise under a nitrogen atmosphere. The resulting mixture was stirred at 0 ° C. for 30 minutes and benzenesulfonyl chloride (0.31 ml, 2.44 mmol) was added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into water (20 ml) and extracted with EtOAc (3 × 10 ml). The combined organic layers were washed with saturated aqueous sodium chloride solution (20 ml), dried over sodium sulfate, filtered, concentrated in vacuo, and 1- (benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3 -B] Pyridine EV-AT6244-001 [607 mg (97.7%)] was obtained as a flesh-colored solid. LCMS (Method D): retention time 1.26 minutes, M / z = 289 (M + 1).

  Ethyl 1- (benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AU3618-002-Step 2

  To a stirred solution of diisopropylamine (0.15 ml, 1.04 mmol) / anhydrous THF (5 ml) at −78 ° C., n-butyl lithium (2.5 M in hexane, 0.37 ml, 0.94 mmol) was added to nitrogen. It was added dropwise over 10 minutes under atmosphere. The resulting mixture was stirred for 30 minutes, warmed to room temperature and stirred for 1 hour. The mixture was then diluted with anhydrous THF (5 ml) and cooled to −30 ° C. 1- (Benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine (EV-AT6244-001, 200 mg, 0.69 mmol) and TMEDA (0.16 ml, 1.04 mmol) / anhydrous THF ( 10 ml) of solution was added via cannula over 20 minutes. The resulting mixture was stirred between −30 ° C. and −20 ° C. for 2.5 hours. Ethyl chloroformate (CAS 541-41-3, 0.20 ml, 2.08 mmol) was added dropwise for 10 minutes and the mixture was stirred at −30 ° C. for 2 hours and allowed to warm to room temperature over 16 hours. . The reaction mixture was cooled to 0 ° C. and quenched with water (15 ml). The aqueous layer was extracted with DCM (3 × 15 ml) and the layers were combined, washed with water (3 × 10 ml), dried over sodium sulfate, filtered and concentrated in vacuo. The crude product product was purified by flash column chromatography (0-100% EtOAc / heptane) to give ethyl 1- (benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2. -Carboxylate EV-AU3618-002 [93 mg (36.5%)] was obtained as an off-white solid. LCMS (Method D): retention time 1.31 min, M / z = 361- (M + 1).

  Note: This reaction was repeated to obtain ethyl 1- (benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylate (165 mg, 29.6%, EV-AU3619-002). Was obtained and combined with EV-AU3618-002 and step 3 was performed.

  Ethyl 6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylate EV-AU3621-002-Step 3

  Ethyl 1- (benzenesulfonyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylate (EV-AU3618-002 and EV-AU3619-002, 84%, 258 mg, 0.60 mmol) To a stirred solution of / THF (10 ml) was added TBAF (1M in THF, 0.78 ml, 0.78 mmol) and the mixture was stirred at 70 ° C. for 30 minutes. The reaction mixture was concentrated to approximately 1/4 volume and purified by flash column chromatography (0-100% EtOAc / heptane) to give ethyl 6-methoxy-1H-pyrrolo [2,3-b]. Pyridine-2-carboxylate EV-AU3621-002 [95 mg (71.7%)] was obtained as an off-white powder. LCMS (Method D): retention time 1.14 minutes, M / z = 221- (M + 1).

1- (3R) -1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3 -Benzodiazole-5-carbonyl} piperidin-3-amine EV-AV3085-001- (EOAI 3449644) I-119

  Methyl 4- (ethylamino) -3-methoxy-5-nitrobenzoate EV-AV3036-001-step 1

  Ethylamine (2M in THF, 18.3 ml) and potassium carbonate (21.1 g, 152.7 mmol) were added to methyl 4-chloro-3-methoxy-5-nitrobenzoate (CAS 63603-09-8, 7.50 g, 30 To a stirred solution of 0.5 mmol) / THF (100 ml). The reaction mixture was stirred at 50 ° C. for 16 hours and then at 60 ° C. for 7 hours. Further potassium carbonate (21.1 g, 152.7 mmol) and ethylamine (2M in THF, 7.63 ml) were added and stirring was continued at room temperature for 60 hours. Volatiles were removed in vacuo and the resulting residue was diluted with EtOAc (150 ml) and washed with water (2 × 50 ml), saturated aqueous sodium chloride solution (50 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to methyl 4- (ethylamino) -3-methoxy-5-nitrobenzoate EV-AV3036-001 [6.88 g (85%)] Was obtained as an orange powder. LCMS (Method D): retention time 1.21 min, M / z = 255 (M + 1).

  Methyl 3-amino-4- (ethylamino) -5-methoxybenzoate EV-AV3040-001-step 2

SnCl ₂ (19.7 g, 103.9 mmol) and 2M HCl (52 ml) were added to methyl 4- (ethylamino) -3-methoxy-5-nitrobenzoate (EV-AV3036-001, 6.88 g, 26.0 mmol). / Ethanol (150 ml) was added to the stirred suspension. The reaction mixture was stirred at reflux for 1 hour and volatiles were removed in vacuo. The resulting residue was basified with 5M aqueous sodium hydroxide (50 ml), then diluted with EtOAc (150 ml) and stirred for 15 minutes. The mixture was then filtered through Kieselguhr and the filtrate was washed with EtOAc. The organic phase of the filtrate was separated and washed with saturated aqueous sodium chloride solution (50 ml). The organic layer was dried over sodium sulfate and concentrated in vacuo to give methyl 3-amino-4- (ethylamino) -5-methoxybenzoate EV-AV3040-001 [4.20 g (71%)] as a brown powder. It was. LCMS (Method D): retention time 0.70 min, M / z = 225 (M + 1).

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxy Rate EV-AV3079-001-step 3

  1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (EV-AR6674-001, 300 mg, 1.36 mmol synthesized according to scheme 1, step 1) / DMF (5 ml) To the solution at 0 ° C. was added HATU (620 mg, 1.63 mmol) and DIPEA (0.47 ml, 2.72 mmol). The mixture was stirred at 0 ° C. for 10 minutes and then 3-amino-4- (ethylamino) -5-methoxybenzoate (EV-AV3040-001, 311 mg, 1.36 mmol) was added. The mixture was heated to 50 ° C. and stirred for 3 hours. The solvent was removed in vacuo and the residue was dissolved in acetic acid (3 ml) and heated in a sealed tube at 80 ° C. for 2 hours. The solvent was removed in vacuo and the crude material was purified by flash chromatography (0-100% EtOAc / heptane) to give methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b. ] Pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxylate EV-AV3079-001 [488 mg (76%)] was obtained as a yellow powder. LCMS (Method D): retention time 1.31 min, M / z = 405 (M + 1).

  2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxylic acid EV-AV3081-001-step 4

  Methyl 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxy The rate (EV-AV3079-001, 488 mg, 1.04 mmol) was dissolved in THF / methanol (4 ml / 1 ml) and 2M aqueous lithium hydroxide solution (5.19 ml) was added. The mixture was stirred at 40 ° C. for 16 hours and the solvent was removed in vacuo. The resulting residue was dissolved in water (2 ml) and acidified to pH 2 using 2M HCl. The mixture was stirred at room temperature for 10 minutes and the precipitate formed was vacuum filtered, dried and 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2- IL] -1-ethyl-7-methoxy-1H-1,3-benzodiazole-5-carboxylic acid EV-AV3081-001 [350 mg (80%)] was obtained as a white powder. LCMS (Method D): retention time 1.14 minutes, M / z = 391- (M + 1).

  Tert-butyl N- (1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3 -Benzodiazole-5-carbonyl} piperidin-3-yl) carbamate EV-AV3083-001-Step 5

  HATU (217 mg, 0.57 mmol) and DIPEA (163 μl, 0.95 mmol) were added to 2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl. To a stirred solution of −7-methoxy-1H-1,3-benzodiazole-5-carboxylic acid (EV-AV3081-001, 200 mg, 0.48 mmol) / DMSO (2 ml) at 0 ° C. The reaction mixture was stirred at 0 ° C. for 10 minutes and then tert-butyl tert-butyl N-[(3R) -piperidin-3-yl] carbamate (CAS 309756-78-3, 100 mg, 0.50 mmol) was added. . The mixture was allowed to warm to room temperature and stirred for 20 minutes. The crude product was purified by preparative HPLC (basic method) and tert-butyl N- (1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine- 2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-yl) carbamate EV-AV3083-001 [211 mg (74%)] as a white powder Obtained. LCMS (Method D): retention time 1.25 min, M / z = 573 (M + 1).

  (3R) -1- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzo Diazole-5-carbonyl} piperidin-3-amine EV-AV3085-001 (EOAI 3449644) -Step 6

  Tert-butyl N- (1- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3 -Benzodiazol-5-carbonyl} piperidin-3-yl) carbamate (EV-AV3083-001, 210 mg, 0.35 mmol) was dissolved in DCM (2 ml) and 2M HCl / diethyl ether (2 ml) was added. It was. The reaction mixture was stirred at room temperature for 2 hours and the solvent was removed in vacuo. The residue was redissolved in water / acetonitrile (2 ml / 0.5 ml), concentrated in vacuo and further dried to give (3R) -1- {2- [1- (cyclopropylmethyl) -1H- Pyrrolo [2,3-b] pyridin-2-yl] -1-ethyl-7-methoxy-1H-1,3-benzodiazol-5-carbonyl} piperidin-3-amine hydrochloride EV-AV3085-001 [ 147 mg (81%)] was obtained as a yellow powder. LCMS (Method A): retention time 2.19 minutes, M / z = 473 (M + 1).

  (1S, 4R, 6S, 7R) -7-Amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-6-ol EV-AW5575-001 (EOAI34559241) I-162 is described in Scheme 10 Tert-butyl N-[(1S, 4R, 6S, 7R) -6-hydroxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate via synthesis of EV-AW5569-001 Synthesized according to the method described in 3:

Scheme 10

  (1S, 4R, 6S, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-Benzodiazole-5-carbonyl} -6-methoxy-2-azabicyclo [2.2.1] heptane-7-amine EV-AW5584-001 (EOAI3359405) I-167 is described in Scheme 10 Tert-Butyl N-[(1S, 4R, 6S, 7R) -6-methoxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate via synthesis of EV-AW5577-001 Synthesis was performed according to the method described in 3.

  (1R, 4R, 6S, 7R) -7-Bromo-6-methoxy-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane EV-AW5568-001-Step 1 -(Scheme 10)

  (4R, 6R) -3-Bromo-1-[(1S) -1-phenylethyl] -1-azatricyclo [2.2.1.0] heptane-1-ium bromide (EV-AW8588-001, 1.00 g , 3.58 mmol) was dissolved in methanol: acetonitrile (1: 1, 40 ml) and the resulting suspension was heated at 65 ° C. for 12 hours. The reaction mixture was concentrated in vacuo and purified by flash column chromatography (0-30% EtOAc / heptane) to give (1R, 4R, 6S, 7R) -7-bromo-6-methoxy-2-[( 1S) -1-Phenylethyl] -2-azabicyclo [2.2.1] heptane EV-AW5568-001 [0.85 g (98%)] was obtained as an orange oil. LCMS (Method D): retention time 0.73 min, M / z = 312 (M + 1).

  (1S, 4R, 6S, 7R) -6-Methoxy-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane-7-amine EV-AW5570-001-Step 2 (Scheme 10)

(1R, 4R, 6S, 7R) -7-Bromo-6-methoxy-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane (EV-AW5568-001,0 To 84 mg, 2.71 mmol) was added 7M ammonia / methanol (7.74 ml). The solution was stirred at 80 ° C. for 2 hours. The reaction mixture was concentrated in vacuo to give (1S, 4R, 6S, 7R) -6-methoxy-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane-7-amine. EV-AW5570-001 [0.82 g (98%)] was obtained as an orange solid.
LCMS (Method D): retention time 0.27 min, M / z = 247 (M + 1).

  Tert-butyl N-[(1S, 4R, 6S, 7R) -6-methoxy-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW5572-001-Step 3 (Scheme 10)

  The title compound is prepared according to the method described in Scheme 10, Step 19, (1S, 4R, 6S, 7R) -6-methoxy-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2. 1] Synthesized from heptane-7-amine (EV-AW5570-001). LCMS (Method D): retention time 0.86 min, M / z = 347 (M + 1).

  Tert-butyl N-[(1S, 4R, 6S, 7R) -6-methoxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW5577-001-Step 4 (Scheme 10)

The title compound is prepared according to the method described in Scheme 10, step 20, tert-butyl N-[(1S, 4R, 6S, 7R) -6-methoxy-2-azabicyclo [2.2.1] heptan-7-yl. It was synthesized from carbamate (EV-AW5572-001).
¹ H NMR (500 MHz, chloroform-d) δ 5.78-5.48 (m, 1H), 3.94 (d, J = 7.6 Hz, 1H), 3.49 (d, J = 6.1 Hz) , 1H), 3.34 (s, 3H), 3.22 (s, 1H), 2.99 (d, J = 9.2 Hz, 1H), 2.47 (d, J = 9.5 Hz, 1H) ), 2.41- (s, 1H), 1.90 (dd, J = 13.5, 7.1 Hz, 1H), 1.75-1.63 (m, 2H), 1.44 (s, 9H). No LCMS data.

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -N-methyl-2-azabicyclo [2.2.1] heptane-7-amine EV-AY4518-001 (EOAI 3462944) I-237 as described in Scheme 10 -Butyl N-[(1R, 4R, 7R) -2-azabicyclo [2.2.1] heptan-7-yl] -N-methylcarbamate described in Scheme 3 via synthesis of EV-AY4514-001 Synthesized according to method.

  (1R, 4R, 7R) -7-Bromo-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane EV-AX4162-002, Step 5 (Scheme 10)

  (4R, 6R) -3-Bromo-1-[(1S) -1-phenylethyl] -1-azatricyclo [2.2.1.0] heptane-1-ium bromide (EV-AW8588-001, 3.10 g , 8.63 mmol) / anhydrous THF (60 ml) was added 4M lithium aluminum hydride / diethyl ether (2.5 ml, 10 mmol) at −10 to −15 ° C. The mixture was stirred at −10 to −15 ° C. for 2 hours and quenched with saturated sodium bicarbonate (60 ml). The mixture was added to water (200 ml) and extracted with ethyl acetate (4 × 100 ml). The extracts were combined, washed with saturated aqueous sodium chloride solution (100 ml), dried over magnesium sulfate, filtered and concentrated in vacuo to give (1R, 4R, 7R) -7-bromo-2-[(1S). -1-Phenylethyl] -2-azabicyclo [2.2.1] heptane EV-AX4162-00 [22.11 g (87%)] was obtained as a brown oil. LCMS (Method D): retention time 0.81 min, M / z = 279.95 / 281.85 (M + 1).

  (1R, 4R, 7R) -2-[(1S) -1-Phenylethyl] -2-azabicyclo [2.2.1] heptane-7-amine EV-AW5552-001-Step 6 (Scheme 10)

  The title compound is prepared according to the method described in Scheme 10, Step 2, (1R, 4R, 7R) -7-bromo-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1. It was synthesized from heptane (EV-AX4162-002). LCMS (Method D): retention time 0.20 min, M / z = 217 (M + 1).

  Tert-butyl N-[(1R, 4R, 7R) -2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW5553-001 Step 9 (Scheme 10)

  The title compound is prepared according to the method described in Scheme 10, Step 19, (1R, 4R, 7R) -2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane-7- Synthesized from amine (EV-AW5552-001). LCMS (Method D): retention time 0.83 min, M / z = 317 (M + 1).

  Tert-butyl N-methyl-N-[(1R, 4R, 7R) -2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV- AY4510-001-Step 10 (Scheme 10)

  tert-Butyl N-[(1R, 4R, 7R) -2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AW5553-001 , 100 mg, 0.32 mmol) / DMF (2 ml) under a nitrogen atmosphere was added sodium hydride (60%, 15 mg, 0.38 mmol) followed by iodomethane (39 μL, 0.63 mmol). The reaction mixture was stirred at room temperature for 16 hours, diluted with water (30 ml) and extracted with EtOAc (2 × 30 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to give tert-butyl N-methyl-N-[(1R, 4R, 7R) -2-[(1S) -1- Phenylethyl] -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AY4510-001 [110 mg (quantitative)] was obtained as a colorless viscous oil. LCMS (Method D): retention time 1.06 minutes, M / z = 331- (M + 1).

  Tert-butyl N-[(1R, 4R, 7R) -2-azabicyclo [2.2.1] heptan-7-yl] -N-methylcarbamate EV-AY4514-001-step 11- (Scheme 10)

The title compound is prepared according to the method described in Scheme 10, Step 20, tert-butyl N-methyl-N-[(1R, 4R, 7R) -2-[(1S) -1-phenylethyl] -2-azabicyclo [2 2.2.1] Heptane-7-yl] carbamate (EV-AY4510-001). ¹ H NMR (500 MHz, methanol-d ₄ ) δ 4.56 (s, 1H), 4.33 (s, 1H), 3.61- (s, 1H), 3.22 (d, J = 10. 9 Hz, 1 H), 2.98 (d, J = 11.1 Hz, 1 H), 2.92-2.87 (m, 3 H), 2.82 (s, 1 H), 1.98-1.83 ( m, 2H), 1.81-1.72 (m, 1H), 1.63-1.56 (m, 1H), 1.48 (s, 9H). No LCMS data.

  (1R, 4R, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1 , 3-Benzodiazole-5-carbonyl} -N, N-dimethyl-2-azabicyclo [2.2.1] heptane-7-amine EV-AY4524-001 (EOAI3468840) I-243 is described in Scheme 10 Of tert-butyl N-[(1R, 4R, 7R) -2-azabicyclo [2.2.1] heptan-7-yl] -N-methylcarbamate EV-AY4523-001 Synthesized according to:

  (1R, 4R, 7R) -N, N-dimethyl-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2.1] heptane-7-amine EV-AY4521-001-step 7 (Scheme 10)

(1R, 4R, 7R) -2-[(1S) -1-Phenylethyl] -2-azabicyclo [2.2.1] heptane-7-amine dihydrochloride (EV-AY4519-001, 210 mg, 0.2 mg). 73 mmol) (synthesized in scheme 10, step 6) / DCM (10 ml) in an aqueous solution of formaldehyde (37%, 0.27 ml, 3.63 mmol) and then STAB (923 mg, 4. 36 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours, diluted with water (30 ml) and extracted with EtOAc (2 × 30 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to give (1R, 4R, 7R) -N, N-dimethyl-2-[(1S) -1-phenylethyl]- 2-Azabicyclo [2.2.1] heptane-7-amine (EV-AY4521-001) [189 mg (quantitative)] was obtained as a colorless viscous oil. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 7.39-7.12 (m, 6H), 3.57 (s, 1H), 3.01- (s, 1H), 2.84 (s, 1H), 2.31-2.12 (m, 3H), 2.07-1.95 (m, 6H), 1.67 (s, 2H), 1.36 (s, 1H), 1.23 −1.12 (m, 3H). No LCMS data.

  Tert-butyl N-[(1R, 4R, 7R) -2-azabicyclo [2.2.1] heptan-7-yl] -N-methylcarbamate EV-AY4523-001-step 8 (Scheme 10)

The title compound is prepared according to the method described in Scheme 10, Step 20, (1R, 4R, 7R) -N, N-dimethyl-2-[(1S) -1-phenylethyl] -2-azabicyclo [2.2. 1] Synthesized from heptane-7-amine (EV-AY4521-001). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 2.41- (s, 1H), 2.12 (dt, J = 9.8, 3.0 Hz, 1H), 1.79 (d, J = 9 0.8 Hz, 1H), 1.49 (s, 1H), 1.47-1.37 (m, 7H), 1.10-0.99 (m, 2H), 0.71-0.61- m, 1H), 0.58-0.48 (m, 1H). No LCMS data.

  3-{[(1S, 4R, 6S, 7R) -7-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7 -Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-6-yl] oxy} propanenitrile EV-AY4932-001 (EOAI3472707) I-256 was converted to tert-butyl N-[(1S, 4R, 6S, 7R) -6- (2-cyanoethoxy) -2- {2- [1- (cyclopropylmethyl) -1H described in Scheme 10 -Pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane -7-yl] carbamate Through the synthesis of EV-AY4931-001, It was synthesized according to the method described in over-time 3. Final deprotection was performed according to the method described in Scheme 1.

  Tert-butyl N-[(1S, 4R, 6S, 7R) -6- (2-cyanoethoxy) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine -2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AY4931- 001-Step 22 (Scheme 10)

  tert-Butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AY5029-001, 120 mg, To a stirred solution of 0.18 mmol) / DMF (3 ml) at 0 ° C. was added sodium hydride (60%, 7.8 mg, 0.20 mmol). The reaction mixture was stirred for 5 minutes and then prop-2-enenitrile (12 μl, 0.18 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed in vacuo and the residue was partitioned between EtOAc (10 ml) and water (10 ml). The aqueous layer was extracted with EtOAc (2 × 5 ml) and the combined organic layers were washed with water (5 ml), dried over sodium sulfate and concentrated in vacuo. The crude product was purified by preparative HPLC (acidic method) and tert-butyl N-[(1S, 4R, 6S, 7R) -6- (2-cyanoethoxy) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [ 2.2.1] Heptane-7-yl] carbamate EV-AY4931-001 [80 mg (67%)] was obtained as a colorless oil. LCMS (Method D): retention time 1.28 minutes, M / z = 640 (M + 1).

  2-{[(1S, 4R, 6S, 7R) -7-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7 -Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-6-yl] oxy} acetonitrile EV-AY4925-001 (EOAI3470051) I -246 was synthesized according to the method described in Scheme 10, Step 22 using bromoacetonitrile. Final deprotection was performed according to the method described in Scheme 1. LCMS (Method A): retention time 2.05 minutes, M / z = 526 (M + 1).

  (1S, 4R, 6E, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-benzodiazole-5-carbonyl} -6- (methoxyimino) -2-azabicyclo [2.2.1] heptane-7-amine EV-AZ4422-001 (EOAI3482317) I-307 N-[(1S, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy- Synthesis of 1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (methoxyimino) -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AZ4420-002 Via the N-Boc deprotection method described in Scheme 3 Synthesized:

  Tert-butyl N-[(1S, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazole-5-carbonyl} -6-oxo-2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AZ4415-001-Step 23 (Scheme 10 )

  tert-Butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AY5029-001, 490 mg, To a solution of 0.84 mmol) / DCM (10 ml) at 0 ° C., desmartin periodinane (710 mg, 1.67 mmol) was added and the reaction mixture was allowed to warm to room temperature. The reaction mixture was stirred for 24 hours, quenched with saturated aqueous sodium thiosulfate solution (10 ml) and extracted with DCM (3 × 20 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The remaining residue was purified by preparative HPLC (acidic method) and tert-butyl N-[(1S, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H- Pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-oxo-2-azabicyclo [2.2. 1] Heptane-7-yl] carbamate EV-AZ4415-001 [471 mg (89%)] was obtained as a white solid. LCMS (Method D): retention time 1.25 min, M / z = 585 (M + 1).

N-[(1S, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl- 1H-1,3-benzodiazole-5-carbonyl} -6- (methoxyimino) -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AZ4420-002-step 24 (Scheme 10 )

  tert-Butyl N-[(1S, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazole-5-carbonyl} -6-oxo-2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AZ4415-001, 50 mg, 0. To a suspension of (08 mmol) / methanol (1 ml) were added O-methylhydroxylamine.HCl (6.7 mg, 0.08 mmol) and sodium bicarbonate (6.6 mg, 0.08 mmol). The reaction mixture is heated at 65 ° C. for 15 hours and then cooled to room temperature. Additional O-methylhydroxylamine.HCl (6.7 mg, 0.08 mmol) and sodium bicarbonate (6.6 mg, 0.08 mmol) were added after 2 hours and the reaction continued at 65 ° C. for an additional 6 hours. . The temperature was lowered to 60 ° C. and the reaction mixture was stirred for 16 hours. The reaction mixture was cooled to room temperature and the solvent was removed in vacuo. The resulting yellow oil was purified by preparative HPLC (acidic method) to give N-[(1S, 4R, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2 , 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (methoxyimino) -2-azabicyclo [2.2. 1] Heptane-7-yl] carbamate EV-AZ4420-002 [12 mg (26%)] was obtained as a white solid. LCMS (Method D): retention time 1.30 minutes, M / z = 614 (M + 1).

  (1R, 4R, 6S, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-6,7-diamine EV-AY5019-001- (EOAI34669927) I-245 was described in Scheme 10 tert-butyl N-[(1R, 4R, 6S, 7R) -6-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] Through the synthesis of -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8585-005 And synthesized according to the method described in Scheme 3.

  Tert-butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl) -2-azabicyclo [2 2.2.1] Heptane-7-yl] carbamate EV-AW8584-001-Step 25 (Scheme 10)

  DIAD (107 μl, 0.51 mmol) was added to a stirred solution of triphenylphosphane (134 mg, 0.51 mmol) / anhydrous THF (5 ml) at 0 ° C. under a nitrogen atmosphere. The reaction mixture was stirred at 0 ° C. for 5 minutes and then tert-butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2, 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [2.2.1] heptane- 7-yl] carbamate (EV-AY5029-001, 200 mg, 0.34 mmol) / anhydrous THF (5 ml) followed by 1H-isoindole-1,3 (2H) -dione (41 μl, 0.34 mmol) It was. The reaction mixture was stirred at room temperature for 18 hours, concentrated in vacuo and purified by flash column chromatography (0-100% EtOAc / heptane) to give tert-butyl N-[(1S, 4R, 6S, 7R). -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole- 5-carbonyl} -6- (1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl) -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8584 -001 [138 mg (48%)] was obtained as a white foam. LCMS (Method D): retention time 1.28 minutes, M / z = 716 (M + 1).

  Tert-butyl N-[(1R, 4R, 6S, 7R) -6-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8585-005-step 26 ( (Scheme 10)

  tert-Butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl) -2-azabicyclo [2 2.2.1] Heptane-7-yl] carbamate (EV-AW8584-001,134 mg, 0.19 mmol) / DCM (3 ml) in a solution of hydrazine hydrate (1: 1) (27 μl, 0.56 mmol) Was added. The reaction mixture was stirred at room temperature for 45 minutes, 50 ° C. for 18 hours, filtered, and the filtrate was concentrated in vacuo. EtOAc (10 ml) was added to the residue and the mixture was stirred for 5 minutes and then filtered. The filtrate was concentrated in vacuo and purified by flash column chromatography (20-100% EtOAc / heptane) to give tert-butyl N-[(1R, 4R, 6S, 7R) -6-amino-2- {2 -[1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl}- 2-Azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8585-005 [75 mg (63%)] was obtained as a white foam. LCMS (Method D): retention time 1.05 min, M / z = 586 (M + 1).

  (1S, 4R, 6S, 7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H -1,3-benzodiazole-5-carbonyl} -N6, N6-dimethyl-2-azabicyclo [2.2.1] heptane-6,7-diamine EV-AW8596-002 (EOAI34776814) I-268, Tert-Butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine-2 as described in Scheme 10 -Yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (dimethylamino) -2-azabicyclo [2.2.1] heptan-7-yl] Through the synthesis of carbamate EV-AW8594-001, It was synthesized according to the method.

  Tert-butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-Methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (dimethylamino) -2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8594-001 Step 27 (Scheme 10)

  tert-butyl N-[(1R, 4R, 6S, 7R) -6-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AW8585-005, 80 mg, To a stirred mixture of 0.12 mmol) / DCM (1 ml) at room temperature was added 3M formaldehyde solution (37% in water, 10 μl). The reaction mixture was stirred for 15 minutes and then sodium tris (acetate-κO) (hydrido) borate (1-) (37 mg, 0.18 mmol) was added. The reaction solution was allowed to continue for 17 hours. Additional sodium tris (acetate-κO) (hydrido) borate (1-) (37 mg, 0.18 mmol) was added and the reaction was continued for 2 hours. The solvent is removed in vacuo and the residue is purified by preparative HPLC (acidic method) to give tert-butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- ( Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6- (dimethylamino) ) -2-Azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8594-001 [20 mg (28%)] was obtained as a white solid. LCMS (Method D): retention time 1.16 minutes, M / z = 614 (M + 1).

  N-[(1S, 4R, 6S, 7R) -7-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7- Methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-6-yl] propanamide EV-AW8592-002 (EOAI34776589) I-264 Tert-butyl N-[(1R, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridine as described in Scheme 10 -2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-propanamido-2-azabicyclo [2.2.1] heptan-7-yl] Through the synthesis of carbamate EV-AW8590-002, scheme 3 Synthesized according to the method described.

  Tert-butyl N-[(1R, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy -1-Methyl-1H-1,3-benzodiazole-5-carbonyl} -6-propanamido-2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AW8590-002-Step 28 (Scheme 10)

  tert-butyl N-[(1R, 4R, 6S, 7R) -6-amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AW8585-005, 62 mg, To a stirred solution of 0.10 mmol) / dioxane (3 ml) was added triethylamine (16 μl, 0.12 mmol) followed by propanoyl chloride (10 μl, 0.12 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated in vacuo and purified by preparative HPLC (acidic method) to give tert-butyl N-[(1R, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl ) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-propanamido-2-azabicyclo [2.2.1] Heptane-7-yl] carbamate EV-AW8590-002 [34 mg (50%)] was obtained as a white solid. LCMS (Method D): 1.19 min, M / z = 642 (M + 1).

(1S, 2R, 8R) -10- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1 , 3-benzodiazole-5-carbonyl} -5-oxa-3,10-diazatricyclo [4.4.0.0 ^{2, 8]} decan-4-one EV-AX4517-002 (EOAI3460130) I- 187 Was synthesized according to the method described in Scheme 10:

  (1S, 4R, 6S, 7R) -7-Amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptan-6-ol EV-AX4510-001-step 29 (Scheme 10)

  The title compound is prepared according to the method described in Scheme 9, Step 6, tert-butyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2 , 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [2.2.1] heptane -7-yl] carbamate Synthesized from EV-AY5029-001. LCMS (Method D): retention time 0.92 min, M / z = 487.15 (M + 1).

  9H-Fluoren-9-ylmethyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-Methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate EV-AX4513-001 -Step 30 (Scheme 10)

  (1S, 4R, 6S, 7R) -7-Amino-2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1 -Methyl-1H-1,3-benzodiazol-5-carbonyl} -2-azabicyclo [2.2.1] heptan-6-ol (EV-AX4510-001, 112 mg, 0.21 mmol) / DCM (2 ml ) Were added DIPEA (0.11 ml, 0.64 mmol) and FMOC chloride (83 mg, 0.32 mmol) at 0 ° C. The resulting mixture was stirred at 0 ° C. for 2 h, saturated aqueous sodium bicarbonate (1 ml) was added and the aqueous layer was extracted with DCM (2 × 1 ml). The combined organic fractions were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by preparative HPLC (acidic method) and 9H-fluoren-9-ylmethyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl ) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [ 2.2.1] Heptane-7-yl] carbamate EV-AX4513-001 [47 mg (31%)] was obtained as a white solid. LCMS (Method D): retention time 1.33 minutes, M / z = 409.4 (M + 1).

(1S, 2R, 8R) -10- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1 , 3-benzodiazole-5-carbonyl} -5-oxa-3,10-diazatricyclo [4.4.0.0 ^{2, 8]} decan-4-EV-AX4517-002- step 31 (scheme 10)

9H-Fluoren-9-ylmethyl N-[(1S, 4R, 6S, 7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-Methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -6-hydroxy-2-azabicyclo [2.2.1] heptan-7-yl] carbamate (EV-AX4513- 001, 46 mg, 0.06 mmol) / DCM (1 ml) at −78 ° C. under a nitrogen atmosphere under N-ethyl-N- (trifluoro-λ˜4-sulfanyl) ethanamine (34 μl, 0 .26 mmol) was added and the reaction solution was stirred at −78 ° C. for 1 h. The reaction solution was warmed to room temperature and stirred for an additional 12 hours. Saturated aqueous sodium hydrogen carbonate (5 ml) was added and the aqueous layer was extracted with DCM (2 × 5 ml). The organic extracts were combined, dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was dissolved in 20% piperidine / DMF (2 ml) and the solution was stirred at room temperature for 2 hours under a nitrogen atmosphere. The reaction mixture was concentrated in vacuo and purified by preparative HPLC (basic method) to give (1S, 2R, 8R) -10- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2 , 3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -5-oxa-3,10-diazatricyclo [4.4.0 .0 ^{2, 8]} decan-4-EV-AX4517-002 [16mg (55% )] as a white solid. LCMS (Method A): retention time 2.90 minutes, M / z = 513.2 (M + 1).

  N-[(7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1, 3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] ethaneimidoamide EV-AZ7933-001 (EOAI3666901) I-340 was described in Scheme 10.1. Synthesized according to method:

N-[(7R) -2- {2- [1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1, 3-Benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] heptan-7-yl] ethaneimidoamide EV-AZ7933-001-Step 1

  (7R) -2- {2- [1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzo Diazole-5-carbonyl} -2-azabicyclo [2.2.1] heptane-7-amine hydrochloride (EV-AU9363-001, 140 mg, 0.28 mmol synthesized according to the method described in Scheme 3) / methanol ( To a stirred solution of 5 ml) was added potassium carbonate (153 mg, 1.10 mmol) and the reaction mixture was stirred at room temperature for 24 hours. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (basic method followed by repurification using acidic method) to give N-[(7R) -2- {2- [1- (cyclopropylmethyl)- 1H-pyrrolo [2,3-b] pyridin-2-yl] -7-methoxy-1-methyl-1H-1,3-benzodiazole-5-carbonyl} -2-azabicyclo [2.2.1] Heptane-7-yl] ethanimidoamidate EV-AZ7933-001 [27 mg (18%)] was obtained as a white powder. LCMS (Method A): retention time 2.05 minutes, M / z = 512.3 (M + 1).

The following compounds were synthesized according to the method described above:

(2- (1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) ( (cis) -Hexahydropyrrolo [3,4-b] [1,4] oxazin-6 (2H) -yl) methanone, 1-349
Note: Starting material was cis-racemic.

Manufactured in the same manner as Scheme 1. Molecular weight = 486.57; LC / MS retention time = 1.19 minutes
(Column: Waters XBridge C18, 2.1 mm x 50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water (containing 0.1% trifluoroacetic acid); mobile phase B: 95: 5 acetonitrile: water ( 0.1% trifluoroacetic acid); Temperature: 50 ° C .; Gradient: 0% B to 100% B over 3 minutes then held at 100% B for 0.75 minutes; Flow rate: 1 mL / min; Detection: MS And UV (220 nm)). M / Z (+) = 487.12; salt = free base.

  (2- (1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) ( (4aR, 7aS) -Hexahydropyrrolo [3,4-b] [1,4] oxazin-6 (2H) -yl) methanone and (2- (1- (cyclopropylmethyl) -1H-pyrrolo [2,4 3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) ((4aS, 7aR) -hexahydropyrrolo [3,4-b] [1 , 4] Oxazin-6 (2H) -yl) methanone to obtain chiral HPLC. Note: Starting material was cis-racemic. Absolute stereochemistry was arbitrarily distributed.

(2- (1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [] produced by the same method as in Scheme 1 d) imidazol-5-yl) ((cis) -hexahydropyrrolo [3,4-b] [1,4] oxazin-6 (2H) -yl) methanone is dissolved in ethanol and then chiral HPLC (Chiralcel OD 21 x 250 mm 10u, wavelength: 254, flow rate: 15 ml / min, solvent A: 100% heptane, solvent B: 100% ethanol, isocratic collection by UV,% B: 35) (1- (Cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) ((4aR, 7aS) -Hexahydropyrrolo [3,4-b] [1,4] oxazine-6 ( 2H) -yl) methanone [25.7 mg (50.4%)] as isomer A and (2- (1- (cyclopropylmethyl) -1H-pyrrolo [2,3-b] pyridin-2-yl) ) -7-Methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) ((4aS, 7aR) -hexahydropyrrolo [3,4-b] [1,4] oxazine-6 (2H) -Yl) methanone [25.4 mg (97.0%)] was obtained as an isomer (absolute stereochemistry assigned arbitrarily).

Isomer A: Chiral purity of I-350 (UV, 254 nm): 100%, retention time: 17.36 minutes (Chiralcel OD 21 x 250 mm 10u, wavelength: 254, flow rate: 15 ml / min, solvent A: 100% heptane, Solvent B: 100% ethanol, isocratic collection by UV,% B: 35). Salt = free base.

  Isomer B: Chiral purity of I-351 (UV, 254 nm): 100%, retention time: 27.17 minutes (Chiralcel OD 21 x 250 mm 10u, wavelength: 254, flow rate: 15 ml / min, solvent A: 100% heptane, Solvent B: 100% ethanol, isocratic collection by UV,% B: 35). Salt = free base.

LC-MS conditions for Schemes 9 and 10:
Column: Waters Acquity SDS
Mobile phase:
Solvent A: Water Solvent B: Acetonitrile Gradient range:
A linear gradient of 2% to 98% solvent B over 1 minute (“min”),
Hold at 98% B for 0.5 minutes ("min").
Gradient time: 1 minute Analysis time: 1.7 minutes Detector: Visualization with ultraviolet light (“UV”) at 254 nanometers (“nm”)

Tert-butyl ((3R, 6S) -1- (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H -Benzo [d] imidazol-5-carbonyl) -6-methylpiperidin-3-yl) carbamate I-352
Prepared using Scheme 11.

  Methyl 2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H-indole-5-carboxylate—Step 2

  1- (Cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (5 g, 20.30 mmol) (prepared from EV-AR3621-002 according to Scheme 2) 1- [Bis (dimethylamino) methylene] -1H-1,2,3-triazolo [4,5-b] pyridinium 3-oxide hexafluorophosphate (HATU, 7.) in dimethylformamide (DMF, 80 mL). 72 g, 20.30 mmol) and N, N-diisopropylethylamine (DIPEA, 3.90 mL, 22.33 mmol). The reaction was stirred for 15 minutes and then methyl 3-amino-4- (methylamino) benzoate EV-AR3155-001 (4.02 g, 22.33 mmol) was added to the reaction solution. The reaction solution was stirred at room temperature for 2 hours. After the reaction was complete, the solvent was removed in vacuo. To the residue was added acetic acid (AcOH, 120 mL) and the vessel was warmed to 70 ° C. and stirred for 3 hours. The solvent was removed and the reaction solution was purified by automated chromatography and the product was eluted from the column with 40% ethyl acetate / hexane to give methyl 2- (1- (cyclopropylmethyl) -6-methoxy. -1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazole-5-carboxylate (6.9 g, 17.67 mmol, 87% yield) was obtained. It was. LCMS: retention time 0.95 min, M / z = 391.0 (M + 1).

  2- (1- (Cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazole-5-carboxylic acid-step 3

  Methyl 2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazole-5-carboxylate ( 500 mg, 1.281 mmol) / THF (7 mL) and methanol (3.50 mL) in water (3.50 mL), lithium hydroxide monohydrate / water solution (3M, 1.281 mL, 3.84 mmol) ) Was added. After 30 minutes, after the starting material precipitated out of solution, additional THF (9 mL) was added and the reaction mixture was stirred overnight. The reaction mixture was concentrated in vacuo and then neutralized with hydrochloric acid (1M in water, 3.84 mL, 3.84 mmol). Water was added to the suspension and the suspension after sonication was filtered and the solid was air dried to give the product 2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2 , 3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazole-5-carboxylic acid (470 mg, 1.249 mmol, 98% yield). LCMS: retention time 0.85 min, M / z = 376.8 (M + 1).

  Tert-butyl ((3R, 6S) -1- (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H -Benzo [d] imidazol-5-carbonyl) -6-methylpiperidin-3-yl) carbamate, I-352-step 4

  2- (1- (Cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazole-5-carboxylic acid (30 mg , 0.080 mmol), tert-butyl ((3R, 6S) -6-methylpiperidin-3-yl) carbamate (25.6 mg, 0.120 mmol), HATU (36.4 mg, 0.096 mmol) and DIPEA (0 0.042 mL, 0.239 mmol) were combined in DMF (1 mL) and the reaction mixture was stirred for 30 min. After amide formation (measured by LCMS) was complete, HCl (4M in dioxane, 1 mL, 4 mmol) was added to the reaction. The reaction was warmed to 50 ° C. and stirred for 3 hours. The reaction mixture was cooled to room temperature, filtered and then purified using preparative HPLC to give ((2S, 5R) -5-amino-2-methylpiperidin-1-yl) (2- (1- (Cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-352 (22 0.6 mg, 0.047 mmol, 59.4% yield, 99% purity). LCMS: retention time 0.73 minutes, M / z = 472.9 (M + 1).

¹ H NMR (500 MHz, DMSO-d6) shift 8.04-8.00 (m, 1H), 7.78-7.72 (m, 2H), 7.40-7.33 (m, 1H), 7.11-7.02 (m, 2H), 6.72-6.63 (m, 1H), 4.54-4.42 (m, 2H), 4.01-3.96 (m, 3H) ), 3.96-3.91 (m, 3H), 3.03-2.91 (m, 1H), 1.95-1.82 (m, 1H), 1.79-1.68 (m) , 2H), 1.66-1.51 (m, 1H), 1.26-1.13 (m, 4H), 0.36-0.29 (m, 2H), 0.25-0.19. (m, 2H).

  ((1R, 4R, 7R) -7-amino-2-azabicyclo [2.2.1] heptan-2-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2 , 3-b] pyridin-2-yl) -1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-353

Following the method of Scheme 11, 1-353 (95% yield, 97% purity) was obtained. LCMS: retention time 0.71 min, M / z = 470.9 (M + 1). (The appearance of some signals was buried under the water peak). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.02 (brd, J = 8.25 Hz, 2H), 7.81- (s, 1H), 7.67-7.78 (m, 1H), 7.42-7.55 (m, 1H), 7.12-7.33 (m, 1H), 7.07 (brs, 2H), 6.68 (d, J = 8.41 Hz, 1H), 4.41-4.59 (m, 2H), 3.97 (brs, 3H), 3.94 (s, 3H), 3.08-3.28 (m, 1H), 2.66 (brs, 1H), 1.79-2.04 (m, 3H), 1.55-1.72 (m, 1H), 1.09-1.28 (m, 1H), 0.31- (brd, J = 7.83 Hz, 2H), 0.22 (brs, 2H).

  ((3S, 5S) -3-Amino-5-fluoropiperidin-1-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2- Yl) -1-methyl-1H-benzo [d] imidazol-5-yl) methanone

Following the method of Scheme 11, I-354 (90% yield, 96% purity) was obtained. LCMS: retention time 0.72 min, M / z = 476.9 (M + 1).
¹ H NMR (500 MHz, DMSO-d6) shift 8.03-7.96 (m, 1H), 7.80-7.70 (m, 2H), 7.44-7.36 (m, 1H), 7.06-7.02 (m, 1H), 6.71-6.60 (m, 1H), 4.56-4.36 (m, 2H), 4.01-3.94 (m, 3H) ), 3.94-3.89 (m, 3H), 3.48-3.33 (m, 1H), 3.07-2.87 (m, 1H), 2.42-2.28 (m , 1H), 1.93-1.72 (m, 1H), 1.21-1.04 (m, 1H), 0.35-0.24 (m, 2H), 0.22-0.08 (m, 2H).

((2S, 5R) -5-amino-2-methylpiperidin-1-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2- Yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I355.
Prepared according to Scheme 12.

Methyl 2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5 -Carboxylate-Step 2
1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid (5 g, 20.30 mmol), HATU (7.72 g, 20.30 mmol) and DIPEA (10 .64 ml, 60.9 mmol) was added to DMF (80 ml). The reaction solution was stirred for 15 minutes at room temperature, at which point methyl 3-amino-5-methoxy-4- (methylamino) benzoate, HCl salt (5.51 g, 22.33 mmol) was added and the reaction solution was added. Stir overnight. The solvent was removed under vacuum and AcOH (120 mL) was added to the residue. The reaction solution was stirred at 70 ° C. for 4 hours, then AcOH was removed in vacuo and the crude product was absorbed on celite and purified using automated chromatography to give methyl 2- (1- (cyclopropyl). Methyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5-carboxylate (6.5 g, 15 .46 mmol, 76% yield). LCMS: retention time 0.98 min, M / z = 421.0 (M + 1).

  2- (1- (Cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5 Carboxylic acid-step 3

  Methyl 2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5 -Carboxylate (500 mg, 1.189 mmol) was dissolved in THF (10 mL), MeOH (5.00 mL) and water (2.500 mL) to give a lithium hydroxide / water solution (3M, 1.189 mL, 3 .57 mmol) was added to the reaction. Additional THF was added until the reaction was homogeneous. The reaction solution was stirred at room temperature for 2 days. The reaction solution was concentrated in vacuo and HCl / water (1M, 3.6 mL, 3.6 mmol) was added to adjust the pH to about 6. The resulting solid was collected and the product 2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1- Methyl-1H-benzo [d] imidazole-5-carboxylic acid (435 mg, 1.07 mmol, 90% yield) was obtained. LCMS: retention time 0.89 min, M / z = 406.8 (M + 1).

  ((2S, 5R) -5-amino-2-methylpiperidin-1-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2- Yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-355-step 4

Following the method of Scheme 12, Step 4, ((2S, 5R) -5-amino-2-methylpiperidin-1-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-355 (68.7% yield, 100% purity) ) Was manufactured. LCMS: retention time 0.76 min, M / z = 502.9 (M + 1).
¹ H NMR (500 MHz, DMSO-d6) shift 8.03-7.98 (m, 1H), 7.28-7.22 (m, 1H), 7.01-6.94 (m, 1H), 6.83-6.77 (m, 1H), 6.70-6.61- (m, 1H), 4.48-4.38 (m, 2H), 4.17-4.06 (m, 3H), 4.00-3.96 (m, 3H), 3.96-3.93 (m, 3H), 2.70-2.62 (m, 1H), 2.55-2.52 ( m, 3H), 1.90-1.88 (m, 2H), 1.76-1.62 (m, 2H), 1.58-1.39 (m, 2H), 1.26-1. 12 (m, 4H), 0.38-0.27 (m, 2H), 0.25-0.14 (m, 2H).

  ((3S, 5S) -3-Amino-5-fluoropiperidin-1-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2,3-b] pyridine-2- Yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-356

Following the method of Scheme 12, ((3S, 5S) -3-amino-5-fluoropiperidin-1-yl) (2- (1- (cyclopropylmethyl) -6-methoxy-1H-pyrrolo [2, 3-b] pyridin-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-356 (56.7% yield, 99% purity) was obtained. It was. LCMS: retention time 0.75 min, M / z = 506.9 (M + 1).
¹ H NMR (500 MHz, DMSO-d6) shift 8.06-7.96 (m, 1H), 7.36-7.25 (m, 1H), 7.01-6.93 (m, 1H), 6.87-6.79 (m, 1H), 6.70-6.61 (m, 1H), 4.44-4.37 (m, 2H), 4.14-4.08 (m, 3H) ), 4.01-3.95 (m, 3H), 3.95-3.92 (m, 3H), 3.54-3.43 (m, 1H), 3.31-3.21 (m) , 1H), 3.06-2.96 (m, 1H), 2.23-2.10 (m, 1H), 1.94-1.84 (m, 2H), 1.65-1.43 (m, 1H), 1.21-1.06 (m, 1H), 0.37-0.24 (m, 2H), 0.23-0.11 (m, 2H).

Tert-butyl ((1S, 4R) -2- (2- (5-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d ] Imidazole-5-carbonyl) -2-azabicyclo [2.2.1] heptan-7-yl) carbamate: 5
Prepared using Scheme 13.1.

5-Bromo-1- (cyclopropylmethyl) -1H-indole-2-carbaldehyde: 2

A cold solution of commercial 5-bromo-1H-indole-2-carbaldehyde 1 (1 g, 4.46 mmol) / DMF (0 ° C.) was treated with NaH (0.196 g, 4.91 mmol) in small portions. . The reaction solution was then stirred at 0 ° C. for a further 30 minutes. (Bromomethyl) cyclopropane (0.723 g, 5.36 mmol) was then added and the ice bath was removed. After 14 hours, the reaction was quenched with H ₂ O (2 mL), then diluted with EtOAc, washed with H ₂ O, brine, dried (MgSO ₄ ) and concentrated to give an orange wax. It was. The wax was then purified by Biotage (80 gcol, 0-60% EtOAc / hexane, 12 CV) to give the desired product [800 mg (64%)] as a pale yellow oil. LCMS (Method A): retention time 2.18 minutes, M / z = 280.10 (M + 2).

Methyl-2- (5-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5-carboxylate: 3

5-Bromo-1- (cyclopropylmethyl) -1H-indole-2-carbaldehyde 2 (2.2 g, 7.91 mmol) and methyl-3-methoxy-4- (methylamino) -5-nitrobenzoate (WO2014) / 015905A1, Intermediate 23) (1.900 g, 7.91 mmol) / ethanol (30 mL) and a solution of sodium hyposulfite (4.13 g, 23.73 mmol) / H ₂ O (10 mL) once. Processed. The reaction solution was then heated at reflux for 18 hours. After this time, the reaction mixture was diluted with DCM (150 ml), dried over MgSO ₄ and concentrated to give an orange solid. The solid was purified by Biotage (80 gcol, 0-30% EtOAc / hexanes, 12 CV) to give recovered aldehyde (1 g) and recovered nitro compound (1.4 g). Obtained with the desired product [430 mg (12%)] as a yellow solid. LCMS (Method A): retention time 2.25 min, M / z = 470.20 (M + 2).

2- (5-Bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5-carboxylic acid: 4

Methyl 2- (5-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5-carboxylate 3 (1.1 g , 2.349 mmol) / THF (15 mL) was treated with a solution of LiOH (0.225 g, 9.39 mmol) / H ₂ O (6 mL) at room temperature. The reaction solution was then heated to reflux for 1.5 hours. The suspension was cooled with ice and then acidified with 1N HCl (ca. 25 ml). The resulting red solid was collected by filtration, washed with H ₂ O and dried (high vacuum). The solid was then taken up in EtOAc / DCM, dried again over MgSO ₄ , filtered and then concentrated to give a red solid [940 mg (88%)]. LCMS (Method A): retention time 1.27 minutes, M / z = 456.20 (M + 2).

tert-Butyl ((1S, 4R) -2- (2- (5-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d ] Imidazole-5-carbonyl) -2-azabicyclo [2.2.1] heptan-7-yl) carbamate: 5

2- (5-Bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazole-5-carboxylic acid 4 (940 mg, A solution of 2.069 mmol), DIEA (0.542 mL, 3.10 mmol) / DCM (50 mL) was treated in one portion with HATU (865 mg, 2.276 mmol). Tert-butyl (1R, 4R, 7R) -2-azabicyclo [2.2.1] heptan-7-ylcarbamate (483 mg, 2.276 mmol) was added and stirring was continued for 18 hours. The reaction mixture was diluted with DCM, washed with H ₂ O, saturated bicarbonate solution, brine, dried (MgSO ₄ ) and concentrated to give an orange oil. The oil was then purified by Biotage (80 g col, 50-90% EtOAc / hexane, 12 CV, then 90% EtOAc / hexane, 2 CV) to give the desired product as an orange oil [1.07 g (80% )]. LCMS (Method A): retention time 1.09 minutes, M / z = 649.50 (M + 1). M / z = 650.13 (M + 2).

Method A

((1S, 4R) -7-amino-2-azabicyclo [2.2.1] heptan-2-yl) (2- (1- (cyclopropylmethyl) -5-((E) -4- (tri Fluoromethyl) styryl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, 1-361:
tert-Butyl ((1S, 4R) -2- (2- (5-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d ] Imidazole-5-carbonyl) -2-azabicyclo [2.2.1] heptan-7-yl) carbamate 5 (50 mg, 0.077 mmol), (E)-(4- (trifluoromethyl) styryl) boronic acid To a suspension of (16.65 mg, 0.077 mmol) and potassium phosphate (III) (0.925 mL, 0.463 mmol) / THF (5 mL) in water (0.5 M) was added 2-dicyclohexylphosphino- 2 ′, 6′-di-I-propoxy-1,1′biphenyl (21.58 mg, 0.046 mmol) and palladium (II) chloride (2.73 mg, 0.015 mmol) were added in one portion. The resulting reddish suspension was heated to 80 ° C. for 4 hours. The reaction solution was diluted with EtOAc, washed with H ₂ O, brine, dried (MgSO ₄ ) and concentrated to give a pale orange wax. This wax was clean enough by LCMS (MW = 739) and proceeded to the deprotection step. The wax was taken up in DCM (2 mL) and then treated with 4M HCl (0.5 mL, 2.000 mmol) (4M in dioxane). After 1 hour, the reaction mixture was concentrated, taken up in DCM, filtered, dried (MgSO ₄ ), concentrated and then purified to give 32 mg in 2 ml DMF. The crude material was purified by preparative LC / MS: column: XBridge C18, 19 × 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water (with 10 mM ammonium acetate); mobile phase B: 95: 5 Acetonitrile: water (with 10 mM ammonium acetate); Gradient: 50-90% B over 19 minutes, then held at 100% B for 4 minutes; Flow rate: 20 mL / min. Fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of product was 20.3 mg (34% yield) and its calculated purity by LCMS analysis was 100%. LCMS (Method B): retention time 2.42 minutes, M / z = 640.14 (M + 1).

Method B

  ((1S, 4R) -7-amino-2-azabicyclo [2.2.1] heptan-2-yl) (2- (5-((2-cyclopropylethyl) amino) -1- (cyclopropylmethyl ) -1H-Indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-yl) methanone, I-374:

A suspension of 5 (50 mg, 0.077 mmol), 2-cyclopropylethanamine (6.56 mg, 0.077 mmol), X-PHOS (7.35 mg, 0.015 mmol) in toluene (15 mL) Degassed with nitrogen for min. Pd ₂ (dba) ₃ (7.06 mg, 7.71 μmol) and sodium tert-butoxide (14.82 mg, 0.154 mmol) were then added and the suspension was heated to 80 ° C. for 4 hours. The reaction solution was cooled and diluted with EtOAc, then washed with H ₂ O, brine, dried (MgSO ₄ ) and concentrated to give a brown wax. LCMS revealed that the wax contained the desired product by LCMS (MW = 652), which was then directly deprotected.

  This material was taken up in DCM (0.5 mL) and then treated with HCl (0.5 mL, 2 mmol, 4M in dioxane). After 1 hour, the reaction was concentrated and then evaporated with DCM (2x). The crude material was dissolved in MeOH (filtered) and evaporated to give a red solid: 37 mg. The crude material was purified by preparative LC / MS: column: XBridge C18, 19 × 200 mm, 5 μm particle size; mobile phase A: 5:95 acetonitrile: water (containing 10 mM ammonium acetate); mobile phase B: 95: 5 Acetonitrile: water (with 10 mM ammonium acetate); Gradient: 35-75% B over 20 minutes, then held at 100% B for 7 minutes; Flow rate: 20 mL / min. Fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of product was 10.9 mg, (16%) and its calculated purity by LCMS analysis was 98%. LCMS (Method B): retention time 1.84 minutes, M / z = 553.26 (M + 1).

Method C

Step 1: tert-butyl ((1S, 4R) -2- (2- (5-cyano-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H- Benzo [d] imidazol-5-carbonyl) -2-azabicyclo [2.2.1] heptan-7-yl) carbamate:

Tert-Butyl ((1S, 4R) -2- (2- (5-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl)-in DMF (4 mL) and water (0.4 mL) 7-methoxy-1-methyl-1H-benzo [d] imidazol-5-carbonyl) -2-azabicyclo [2.2.1] heptan-7-yl) carbamate (100 mg, 0.154 mmol) 5 , zinc cyanide A suspension of (18.10 mg, 0.154 mmol), Pd2dba3 (7.06 mg, 7.71 μmol) and DPPF (10.26 mg, 0.019 mmol) was heated to 120 ° C. for 18 hours. Additional zinc cyanide (18.10 mg, 0.154 mmol), Pd2dba3 (7.06 mg, 7.71 μmol) and DPPF (10.26 mg, 0.019 mmol) were added and stirring was continued at 120 ° C. for 4 hours. The reaction solution was cooled, diluted with EtOAc, washed with H ₂ O, saturated bicarbonate solution, dried (MgSO ₄ ) and concentrated to give a black wax. The wax was purified by Biotage (40 gcol, 20-100% EtOAc / hexane, 12 CV, then 100% EtOAc for 4 CV) to give the desired product as a pale yellow wax: 89 mg (97% yield) LCMS (Method A): Retention time 2.11 min, M / z = 595.65 (M + 1).

Step 2: 2- (5-((1S, 4R) -7-amino-2-azabicyclo [2.2.1] heptane-2-carbonyl) -7-methoxy-1-methyl-1H-benzo [d] Imidazole-2-yl) -1- (cyclopropylmethyl) -1H-indole-5-carbonitrile, I-370:

  tert-Butyl ((1S, 4R) -2- (2- (5-cyano-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d ] Imidazole-5-carbonyl) -2-azabicyclo [2.2.1] heptan-7-yl) carbamate (35 mg, 0.059 mmol) / DCM (1 mL) was added to HCl (0.5 ml, 2 mmol, dioxane). In 4M). After 1 hour, the reaction was concentrated and then dissolved sequentially in DCM and concentrated (2x). The crude material was dissolved in MeOH (filtered) and evaporated to give a red solid (22 mg) that was dissolved in DMF (1.5 ml) and purified by preparative LC / MS: Column: XBridge C18, 19 × 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water (containing 10 mM ammonium acetate); mobile phase B: 95: 5 acetonitrile: water (containing 10 mM ammonium acetate); gradient : 15-55% B over 20 minutes, then hold at 100% B for 5 minutes; flow rate: 20 mL / min. Fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of product was 14.4 mg (38% yield) and its calculated purity by LCMS analysis was 100%. LCMS (Method B): retention time 1.81 min, M / z = 495.07 (M + 1).

Tert-butyl ((1S, 4R) -2- (2- (6-bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d ] Imidazole-5-carbonyl) -2-azabicyclo [2.2.1] heptane-7-yl) carbamate 6 :

  This bromide was prepared as described in Scheme 13.1, starting from commercially available 6-bromo-1H-indole-2-carbaldehyde to obtain an orange wax. LCMS (Method A): retention time 1.29 minutes, M / z = 650.30 (M + 2).

All six substituted -1H-indol-2-yl compounds were converted to (Scheme 13), tert-butyl ((1S, 4R) -2-, using the same method as described for 5-substituted compounds. (2- (6-Bromo-1- (cyclopropylmethyl) -1H-indol-2-yl) -7-methoxy-1-methyl-1H-benzo [d] imidazol-5-carbonyl) -2-azabicyclo [ 2.2.1] Heptane-7-yl) carbamate 6 .

HPLC Method Method A for Schemes 13 and 14
(Low pH Shimadzu 3 minutes method)
Column: Waters Acquity UPLC BEH dC18, 2.1 mm x 50 mm, 1.7 μm Column Flow rate: 0.6 ml / min Mobile phase:
A, 0.05% TFA (aqueous solution) and B, 0.05% TFA (acetonitrile)
Injection volume: 3 μl
Temperature: 40 ° C
Detection: 220 nm (nominal)
Gradient-0-100% B

Method B
Column: Water Xbridge C18, 2.1 mm x 50 mm, 1.7 μm Column Flow rate: 0.6 ml / min Mobile phase:
Acetonitrile: water (containing 10 mM ammonium acetate);
Mobile phase B:
95: 5 acetonitrile: water (containing 10 mM ammonium acetate);
Temperature: 50 ° C .;
Gradient: 0% B to 100% B over 3 minutes, then hold at 100% B for 0.75 minutes Injection volume: 3 μl
Temperature: 40 ° C
Detection: 220 nm (nominal)

The following compounds were prepared using Method A, B or C as described in Schemes 13 and 14:

Biological Assays Compounds of the invention were assayed as inhibitors of PAD4 using the following assay protocol.

Compounds are dissolved in 100% DMSO to reach a final concentration of 100 mM compound. Compound stock solutions were stored at RT. Serial dilutions were prepared in DMSO and mixed 8 times with a 20 μL mixing volume. The final assay conditions are as follows:
Reaction volume: 20 μl
Assay buffer (as above): 100 mM Tris-HCl (pH 7.6), 2 mM DTT, 1 mM CaCl ₂
Final concentration:
-100 nm HPAD4 enzyme -50MyuM (hereinafter eight times K _m) substrate peptide -0.5% DMSO
Total incubation time: 65 minutes at 37 ° C. Stop solution: 40 μl 5% TCA / ACN
Compound solution (0.25 μL) was added to 200 nM PAD4 (10 μL) in assay buffer (100 mM Tris-HCl (pH 7.6), 2 mM DTT). After 5 minutes, 100 μM substrate (10 μL) in buffer (100 mM Tris-HCl (pH 7.6), 2 mM DTT, 2 mM CaCl ₂ ) was added and the reaction was incubated for 60 minutes at 37 ° C. The enzyme reaction was quenched by adding 5% TCA / ACN (1.7% TCA final concentration) stop solution (40 μl). A substrate containing arginine and a product containing citrulline (+1 Da mass shift) are subjected to a solid phase extract on the Agilent RapidFire (RF) 300 System for multiple reaction monitoring (MRM) for quantification. And detected with a double pole, triple pole, quadrupole Agilent 6460 QQQ mass spectrum analyzer (MS) instrument.

The IC ₅₀ for determining a given test compound IC ₅₀ for PAD isozymes, as measured using a mass spectrum assay, which was detected citrullinated product BAEE after the reaction with phenylglyoxal. Test compounds were dissolved in 100% DMSO and 0.125 uL was transferred to a 384 well REMP polypropylene plate prior to enzyme addition. Enzymes and compounds are preincubated for 30 minutes at 37 ° C. and BAEE substrate is added such that the final concentration in each assay well is equal to the Km value for BAEE at the time when the calcium concentration is saturated ( 250 uM), the reaction was started. The buffer used for this reaction was 25 mM Hepes (pH 7.5), 5 mM NaCl, 1 mM DTT, 0.01% Chaps, 0.2 mg / mL BSA, and either 50 uM or 1 mM CaCl ₂ (this is BAEE when measured at 10 times Km, each containing corresponds to one fifth of 20XK _0.5 for _{K 0.5} or ^{Ca 2+} for ^{Ca 2+).} The typical enzyme concentration used was 5 nM in the final reaction and the total reaction volume was 25 uL. The reaction was carried out at 37 ° C. for 1.5 hours after the addition of BAEE and then quenched with 15 ul 6.1 N TCA and 35 ul 8.5 mM phenylglyoxal. The final concentration of phenylglyoxal was 4 mM. The mixture was incubated with stirring for an additional 30 minutes at 37 ° C. to complete the modification of the citrullinated product with phenylglyoxal. The quenched reaction plate was centrifuged at 5000 × g for 3 minutes and an equal volume of methanol containing an internal standard (phenylglyoxal modified citrulline) was added to each well. The reaction was transferred to a new 384 well REMP plate for Rapidfire MS analysis.

  Samples are layered on a RapidFire RF300 System (Agilent) (in which the samples are first transported during 1000 ms) and then directly onto a C18 separation cartridge using a mixture of acetonitrile containing 0.01% formic acid. Layered and desalted for 3000 ms. The mobile phase flow rate was 1.5 ml / min. Once the sample elutes from the cartridge, the sample is transferred into the mass spectrometer for 4000 ms at a flow rate of 1.25 ml / min using a mobile phase of acetonitrile containing 0.01% formic acid. Peptidyl citrulline and internal standard ions were analyzed using a Sciex API4000 triple quadrupole mass spectrometer (Applied Biosystems) equipped with ESI. Product and internal standard MRM transitions were monitored at m / z 424.5-30.4 and m / z 293-247, respectively. The data measurement time (dwell time) for each transition was set to 200 ms and the ESI voltage was used at 5500 V at a source temperature of 400 ° C. Differentiation was performed using the extracted ion peak for each transition. Peak areas were normalized to the internal standard.

For determination of IC50, compounds were tested at 11 different concentrations, serially diluted 3-fold with DMSO. The percentage of peak area was calculated by dividing the peak area of the analyte by the peak area of the internal standard. The ratio of the peak area of DMSO control to enzyme-free background was used to calculate as% inhibition occurring at each concentration of inhibitor, and IC ₅₀ was calculated using the following equation:

Where Y =% inhibition at each inhibitor concentration, A = minimum Y value, B = maximum Y value, C = log IC ₅₀ , D = hill slope, and x = inhibitor concentration.

Table 2 below shows the activity of selected compounds of the present invention in the PAD4 assay described above. The compound numbers correspond to the compound numbers in Table 1. Compounds exhibiting activity represented as “A” provide an IC ₅₀ of < 1 μM; compounds exhibiting activity represented as “B” provide an IC ₅₀ of 1.0-5.0 μM; Compounds exhibiting activity represented as “C” provide an IC ₅₀ of 5.0 to 10.0 μM; and compounds exhibiting activity represented as “D” provide an IC ₅₀ of > 10.0 μM. . The term pIC ₅₀ = −log (IC ₅₀ ).
Compounds exhibiting activity represented as “E” exhibit a pIC ₅₀ < 4; compounds exhibiting activity represented as “F” provide a pIC ₅₀ of 4.0-5.0; “G” Compounds exhibiting activity expressed as provide a pIC ₅₀ of 5.0-6.0; compounds exhibiting activity expressed as “H” provide a pIC ₅₀ of > 6. “NA” represents “not assayed”.

Table 2. PAD4 activity

Claims (21)

Formula I ′:

[Where
Ring A is



Because
Ring A is optionally substituted with 1 to 4 groups selected from fluorine, —CN, —OR or C _1-6 aliphatic groups (optionally substituted with 1 to 3 fluorine atoms). May have been;
Ring B is

Is;
R ¹ is hydrogen, —CN, —OR,
Or a C _1-6 aliphatic group (optionally substituted with 1 to 4 groups selected from fluorine, —CN or —OR);
R ² is hydrogen or a C _1-10 aliphatic group optionally substituted with 1 to 5 groups selected from fluorine, —CN or —OR;
Each of X ¹ and X ² is independently selected from N or C (R ⁴ );
R ³ is halogen, —CN, —R,

Or -OR;
Each R ⁴ is independently halogen, —CN, —R,
Is;
R ⁵ is hydrogen or halogen;
n is 0-4; and each R is independently hydrogen or a C _1-6 aliphatic group optionally substituted with 1 to 3 fluorine atoms.
Or a pharmaceutically acceptable salt thereof. Ring A is
The compound of claim 1, wherein Ring A is
The compound of claim 1, wherein The compound according to any one of claims 1 to 3, wherein R ¹ is methyl, ethyl or propyl. The compound according to any one of claims 1 to 4, wherein R ² is a C _1-10 aliphatic group which may be optionally substituted with 1 to 5 fluorine atoms. R ² is methyl, ethyl or -CH ₂ - cyclopropyl, 6. The compound of claim 5, wherein. R ² is 1-5 _{C 1-10} aliphatic group substituted with a fluorine atom, 6. The compound of claim 5, wherein. R ² is _-CH 2 CF _3, claim 7 compound according. Both of X ¹ and ^{X 2} is N, compounds of any one of claims 1-8. The compound according to any one of claims 1 to 8, wherein X ¹ is N and X ² is CH. The compound according to any one of claims 1 to 8, wherein X ¹ is CH and X ² is N. The compound according to any one of claims 1 to 8, wherein both X ¹ and X ² are CH. The compound according to any one of claims 1 to 12, wherein R ⁴ is a C _1-6 aliphatic group or -OR. R ⁴ is ethyl or -OCH _3, claim 13 A compound according.   A pharmaceutically acceptable composition comprising a compound according to any one of claims 1 to 14 and a pharmaceutically acceptable carrier, adjuvant or vehicle.   16. A composition according to claim 15 in combination with another therapeutic agent.   A method of inhibiting PAD4 in a subject or biological sample, comprising contacting PAD4 with a compound according to any one of claims 1-14.   16. A method of treating a PAD4-mediated disease, disorder or symptom, comprising administering to the subject the composition of claim 15.   The method of claim 18, wherein the subject is a human subject.   PAD4-mediated diseases, disorders or symptoms include acid-induced lung injury, acne (PAPA), acute lymphoblastic leukemia, acute respiratory distress syndrome, Addison's disease, adrenal hyperplasia, adrenal cortical dysfunction, aging, AIDS, alcoholic Hepatitis, alcoholic hepatitis, alcoholic liver disease, allergic asthma, allergic bronchopulmonary aspergillosis, allergic conjunctivitis, alopecia, Alzheimer's disease, amyloidosis, amyotrophic lateral sclerosis and weight loss, angina, blood vessels Edema, sweatless ectodermal dysplasia (AHED-ID), ankylosing spondylitis, anterior ocular inflammation, antiphospholipid syndrome, aphthous stomatitis, appendicitis, arthritis, asthma, atherosclerosis, atopic Skin inflammation Bone pain, bronchiolitis, burns, bursitis, cancer, cardiac hypertrophy, carpal tunnel syndrome, cataract disorder, cataract, cerebral aneurysm, chemical irritant-induced inflammation, chorioretinitis, chronic heart disease, Chronic lung disease in premature infants, chronic lymphocytic leukemia, chronic obstructive pulmonary disease, colitis, complex local pain syndrome, collagen disease, corneal ulcer, Crohn's disease, cryopyrin-related periodic fever syndrome, cryptococcosis, cystic fibrosis, Interleukin-1-receptor antagonist (DIRA) deficiency, dermatitis, dermatitis endotoxemia, dermatomyositis, childhood brain stem glioma, endometriosis, endotoxemia, epicondylitis, erythrocytopenia Disease, familial amyloid polyneuropathy, familial cold urticaria, familial Mediterranean fever, fetal growth retardation, glaucoma, glomerular disease, glomerulonephritis, gout, gouty arthritis, graft-versus-host disease, gastrointestinal disease Head injury, headache, hearing loss, heart disease Hemolytic anemia, Henoch-Schöhlein purpura, hepatitis, hereditary autoinflammatory syndrome, herpes zoster and herpes simplex, HIV-1, Hodgkin's disease, Huntington's disease, pulmonary hyaline, hyperammonemia, hypercalcemia, Hypercholesterolemia, hyperimmunoglobulinemia D syndrome with periodic fever (HIDS), aplastic anemia and other anemias, aplastic anemia, idiopathic thrombocytopenic purpura, dyschromia, infectious mononuclear Sphere disease, inflammatory bowel disease, inflammatory lung disease, inflammatory neuropathy, inflammatory pain, insect bite-induced inflammation, iritis, irritation-induced inflammation, ischemia / reperfusion, juvenile rheumatoid arthritis, keratitis , Kidney disease, kidney damage caused by parasitic infection, kidney damage causing parasitic infection, prevention of kidney transplant rejection, leptospirosis, leukemia, Lefler syndrome, lung trauma, lung trauma, lupus, lupus, lupus nephritis, lymphoma , Meningitis, mesothelioma, mixed collagen disease, Maccle Wells syndrome (Hearing loss amyloidosis), multiple sclerosis, muscle fatigue, muscular dystrophy, myasthenia gravis, myocarditis, mycosis fungoides, mycosis fungoides Tumor, myelodysplastic syndrome, myositis, rhinosinusitis, necrotizing enterocolitis, neonatal-onset multi-organ inflammatory disease (NOMID), nephrotic syndrome, neuritis, neuropathological disease, non-allergy-induced asthma , Obesity, ocular allergy, optic neuritis, organ transplantation, osteoarthritis, otitis media, Paget's disease, pain, pancreatitis, Parkinson's disease, pemphigus, pericarditis, periodic fever, periodontitis, peritoneal endometriosis, pertussis Pharyngitis and lymphadenitis (PFAPA syndrome), plant stimulus-induced inflammation, pneumonia, pneumonitis, pneumonitis, pneumoniasis / urushiol oil-induced inflammation, polyarteritis nodosa, polychondritis Frequent Cystic kidney disease, polymyositis, psoriasis, psoriasis, psoriasis, psoriasis, psychosocial stress disease, lung disease, pulmonary hypertension, pulmonary fibrosis, pyoderma gangrenosum, pyogenic arthritis, kidney disease, retinal disease, rheumatism Congenital carditis, rheumatic disease, rheumatoid arthritis, sarcoidosis, seborrhea, sepsis, severe pain, sickle cell, sickle cell anemia, silica-induced disease, Sjogren's syndrome, skin disease, sleep apnea, solid tumor, Spinal cord injury, Stevens-Johnson syndrome, stroke, subarachnoid hemorrhage, sunburn, temporal arteritis, tendon synovitis, thrombocytopenia, thyroiditis, tissue transplantation, TNF receptor-related periodic syndrome (TRAPS), toxoplasma Selected from the group consisting of infectious disease, transplantation, traumatic brain injury, tuberculosis, type I diabetes, type II diabetes, ulcerative colitis, urticaria uveitis, uveitis and Wegener's granulomatosis 18 method described.   19. The method of claim 18, wherein the PAD4-mediated disease, disorder or condition is selected from rheumatoid arthritis, vasculitis, systemic lupus erythematosus, ulcerative colitis, cancer, cystic fibrosis, asthma, cutaneous lupus erythematosus and psoriasis. .